Академический Документы
Профессиональный Документы
Культура Документы
Analysis
1st Edition
M A N U A L
Authors:
Robert Gates
Elner Rathbone
Lisa Masterson
Ian Wright
Asgar Electricwala
Glycan
Classification
Glycoprotein
Purification
Deglycosylation
Strategies
Detection
Glycan Analysis
Product Directory
sigma-aldrich.com
Glycoprotein Analysis
Application Note
PNGase F Enzyme:
Efficient N-Deglycosylation
in Glycoprotein Analysis
By Elner Rathbone, Asgar Electricwala, and Ian Wright
Sigma-Aldrich Corporation, Poole, England
Introduction
Glycosylation is one of the most common post-translational
modifications of proteins in eukaryotic cells. These glycoproteins are involved in a wide range of biological functions such as receptor binding, cell signaling, immune
recognition, inflammation, and pathogenicity. Mammalian
glycoproteins contain three major types of oligosaccharides (glycans): N-linked, O-linked, and glycosylphosphatidylinositol (GPI) lipid anchors. N-Linked glycans are attached
to the protein backbone via an amide bond to an asparagine
residue in an Asn-Xaa-Ser/Thr motif, where X can be any
amino acid, except Pro. O-Linked glycans are linked via the
hydroxyl group of serine or threonine. Variation in the
degrees of saturation at available glycosylation sites results
in heterogeneity in the mass and charge of glycoproteins.
To study the structure and function of a glycoprotein, it is
often desirable to remove all or just a select class of glycans.
This approach allows the assignment of specific biological
functions to particular components of the glycoprotein.
The removal of N-linked glycans from glycoproteins eliminates heterogeneity in matrix-assisted laser desorption/
ionization-time of flight (MALDI-TOF) MS analysis. Also,
removal of glycans may enhance or reduce the blood
clearance rate and/or potency of a glycoprotein therapeutic. Although sites of potential N-glycosylation can be
predicted from the consensus sequence Asn-Xaa-Ser/Thr,
it cannot be assumed that a site will actually be glycosylated. Therefore the sites of glycan attachment must be
identified and characterized by analytical procedures.
Peptide-N-glycosidase F (PNGase F) is one of the most widely
used enzymes for the deglycosylation of glycoproteins.
The enzyme releases asparagine-linked (N-linked) oligosaccharides from glycoproteins and glycopeptides. A tripeptide
with the glycan-linked asparagine as the central residue
is the minimum substrate for PNGase F. The glycan can
be a high-mannose, hybrid, or complex type. However,
N-glycans with fucose linked a(1,3) to the Asn-bound
N-acetylglucosamine are resistant to the action of PNGase F.
MALDI-TOF MS is a widely used technique for rapid identification of proteins separated by gel electrophoresis.
Glycopeptides, however, suffer from signal suppression
Table of Contents
Overview
Key to Monosaccharide Symbols,
Abbreviations and Projections . . . . . . . . . . . . 02
Classification and Structure of
Glycan Components
N-Linked Glycans . . . . . . . . . . . . . . . . . . . 05
O-Linked Glycans . . . . . . . . . . . . . . . . . . . 09
GPI Anchored Proteins . . . . . . . . . . . . . . . 12
Glycoprotein Purification
m-Aminophenylboronate Matrices . . . . . . . . 13
Lectin Matrices . . . . . . . . . . . . . . . . . . . . . . . . 13
Glycoprotein Detection
Colorimetric Detection on PAGE and Blots .
Fluorescent Detection on PAGE . . . . . . . . .
Biotin Labeling on Blots . . . . . . . . . . . . . . .
Glycoprotein Standards . . . . . . . . . . . . . . .
.
.
.
.
.
.
.
.
15
16
17
18
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
24
25
29
32
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
36
37
39
45
45
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
47
49
49
55
57
60
63
65
67
67
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
. 68
. 69
. 74
. 74
. 75
. 76
. 79
. 80
Overview
One of the distinguishing features of the
proteome in eukaryotic cells is that most
proteins are subject to post-translational
modification, of which glycosylation is the
most common form. It is estimated that
more than half of all proteins that have
been characterized are glycoproteins. The
carbohydrate components of glycoproteins
perform critical biological functions in
protein sorting, immune and receptor
recognition, inflammation, pathogenicity,
metastasis, and other cellular processes.
Mammalian glycoproteins contain three
major types of oligosaccharides (glycans):
N-linked, O-linked, and glycosylphosphatidylinositol (GPI) lipid anchors. N-Linked glycans
are linked to the protein backbone via an
amide bond to asparagine residues in an
Asn-X-Ser/Thr motif, where X can be any
amino acid, except Pro. O-Linked glycans
are linked to the hydroxyl group of serine
Haworth projection
OH
HO
CH2OH
O
OH
HO
OH
HO
OH
Fischer projection
H
HO
OH
OH
OH
Glc
OH
CH2OH
b-D-Glucose (Glc)
OH
sigma-aldrich.com
OH
CH2OH
HO
HO
HO
b-D-Mannose (Man)
Symbol
OH
OH
OH
OH
HO
HO
HO
OH
OH
H
CH2OH
Man
Haworth projection
HO
OH
CH2OH
HO
OH
Symbol
H
O
OH
OH
OH
OH
HO
Fischer projection
HO
HO
Gal
OH
H
OH
CH2OH
b-D-Galactose (Gal)
OH
CH2OH
O
HO
OH
HO
HO
OH
NHAc
OH
NHAc
HO
OH
H
NHAc
GlcNAc
OH
H
CH2OH
b-D-N-Acetylglucosamine (GlcNAc)
CH2OH
OH
HO
OH
OH
HO
H
OH
HO
OH
NHAc
NHAc
HO
HO
GalNAc
NHAc
b-D-N-Acetylgalactosamine
(GalNAc)
CH2OH
HO
HO
OH
OH
OH
b-D-Xylose (Xyl)
H
HO
OH
OH
OH
OH
OH
Xyl
H
H
sigma-aldrich.com
O
HO
Haworth projection
Fischer projection
CO2H
HO
CH2OH
OH
HO
CO2H
AcHN HO
AcHN
OH
OH
OH
CO2H
OH
OH
HO
OH
a-N-Acetylneuraminic acid
Sialic Acid (NeuNAc)
OH
OH
CH2OH
CH2OH
CO2H
HO
O
OH
OH
O
HO
OH
OH
HO
HO
OH
OH
H
OH
H
CO2H
HO
O
OH
HO2C
OH
HO
OH
OH
IdoA
OH
OH
H
CO2H
OH
H3C
HO
sigma-aldrich.com
OH
a-L-Fucose (Fuc)
CH2OH
OH
HO
HO
GlcA
OH
HO
NeuNAc
AcHN
OH
Symbol
CH3
O
OH
OH
HO
HO
OH
OH
OH
OH
OH
H
CH3
Fuc
sigma-aldrich.com
sigma-aldrich.com
High-Mannose Structure
sigma-aldrich.com
Hybrid Structure
sigma-aldrich.com
O-Linked Glycans
O-Linked glycans are usually attached to the peptide chain
through serine or threonine residues. O-Linked glycosylation is a true post-translational event and does not require
a consensus sequence and no oligosaccharide precursor
is required for protein transfer. The most common type
of O-linked glycans contain an initial GalNAc residue (or
Tn epitope), these are commonly referred to as mucin-type
glycans. Other O-linked glycans include glucosamine, xylose,
galactose, fucose, or manose as the initial sugar bound to
the Ser/Thr residues. O-Linked glycoproteins are usually
large proteins (>200 kDa) that are commonly bianttennary
with comparatively less branching than N-glycans.
Glycosylation generally occurs in high-density clusters
and may contribute as much as 50-80% to the overall mass.
O-Linked glycans tend to be very heterogeneous, hence
they are generally classified by their core structure. Nonelongated O-GlcNAc groups have been recently shown to
be related to phosphorylation states and dynamic processing
sigma-aldrich.com
10
Core 1
Core 2
Core 3
sigma-aldrich.com
Core 4
11
Core 5
Core 6
Core 7
sigma-aldrich.com
Core 8
12
sigma-aldrich.com
GPI Anchor
13
Glycoprotein Purification
Glycoprotein Purification
Procedure
Equilibration buffers should be of low ionic strength,
with pH 7-9.
For a column volume of 1 ml, apply 1-2 mg of protein in
approximately 250 ml of buffer: 50 mM taurine/NaOH,
pH 8.7, containing 20 mM MgCl2.
Optimize the column flow rate to 2 ml/hour, collecting
2 ml fractions.
Elute the bound protein using the same buffer with
50 mM sorbitol or 50 mM Tris/HCl added.
References
1. Mallia, A.K. et al., Anal. Letters, 14 (B8), 649-661 (1981).
2. Immobilized Affinity Ligand Techniques, Hermanson, G.T., et al.,
Eds. (Academic Press, 1992), pp. 338, 339-392.
3. Affinity Chromatography: A Practical Approach, P.D.G Dean, W.S.
Johnson and F. A. Middle, Eds., (IRL Press, 1985), p. 133.
Aminophenylboronate Matrices
A 8530
m-Aminophenylboronic Acid
Affinity Medium
A 4046
m-Aminophenylboronic Acid
Affinity Medium
A 8312
m-Aminophenylboronic Acid
Affinity Medium
Elution:
Elute the target protein with gradient or step-wise
elution with methyl a-D-glucopyranoside or methyl
a-D-mannopyranoside, glucose, or mannose (5 mM 500 mM).
Maximum recovery and cleaning of the resin may be
achieved by using 1 M sucrose, glucose, mannose, or
corresponding a-methyl glycoside. The addition of
chaotropic agents (0.5 M to 6 M) may also be required
for maximum recovery, but these denaturing conditions
may severely damage the resin. Therefore they should
only be used as a last resort.
sigma-aldrich.com
Lectin Matrices
14
Glycoprotein Purification
Glycoprotein Purification
Con A Matrices
C 9017
Concanavalin A Immobilized
Matrix: Sepharose 4B
Activation: cyanogen bromide
Ligand immobilized: 10-15 mg per ml
Binding Capacity: 20-45 mg thyroglobulin per ml
Form: suspension in 0.1 M acetate buffer, pH 6.0, containing 1 M NaCl, 1 mM each of
CaCl2, MgCl2, and MnCl2 and 0.01% thimerosal
C 6170
Concanavalin A Immobilized
References
1. Yahara, I. and Edelman, G.M., Restriction of the mobility of lymphocyte immunoglobulin receptors by concanavalin A. Proc. Nat.
Acad. Sci. USA, 69(3), 608-612 (1972).
2. Li, Y., et al., The p185neu-containing Glycoprotein Complex of a
Microfilament-associated Signal Transduction Particle. J. Biol.
Chem., 274(36), 25651-25658 (1999).
Elution:
Elute the target protein with gradient or step-wise
elution with equilibration buffer containing 100 mg/ml
N-acetylglucosamine (Product Code A 8625).
Procedure
Equilibration and Binding:
Wash and equillibrate column with 5 column volumes
of wash solution (0.05 M sodium phosphate, pH 7.0,
containing 0.2 M NaCl).
Load sample solution in equilibration buffer (protein
concentrations 1-20 mg/ml, free of particulates).
Wash the resin with equilibration buffer until eluent is
protein free.
WGA Matrices
L 1882
L 1394
References
sigma-aldrich.com
1. Lotan, R., et al., Activities of lectins and their immobilized derivatives in detergent solutions. Implications on the use of lectin
affinity chromatography for the purification of membrane glycoproteins. Biochem., 16, 1787-1794 (1977).
2. Janicott, M., et al., The insulin-like growth factor 1 (IGF-1) receptor is responsible for mediating the effects of insulin, IGF-1, and
IGF-2 in Xenopus laevis oocytes. J. Biol. Chem., 266, 9382 (1991).
15
Glycoprotein Detection
Glycoprotein Detection
Steps
1. Fixing
30 min.
60 min.
2. Washing
2 x 10 min.
2 x 20 min.
3. Oxidation
30 min.
60 min.
4. Washing
2 x 10 min.
2 x 20 min.
5. Staining
6. Reduction
60 min.
120 min.
7. Washing
8. Storage
overnight
overnight
sigma-aldrich.com
16
Glycoprotein Detection
Fluorescent Detection on PAGE Gels
Glycoprotein Detection
GlycoProfile III
Fluorescent Glycoprotein Detection Kit
8 Product Code PP0300
Product Description
GlycoProfile III is designed for the fluorescent in-gel
detection of glycosylated proteins utilizing standard
UV-transillumination. Following SDS-PAGE, proteins are
fixed in the gel with an acetic acid/methanol solution.
The carbohydrates on the proteins are oxidized to aldehydes with periodic acid. A hydrazide dye is reacted with
the aldehydes, forming a stable fluorescent conjugate.
This allows for the specific, sensitive detection of the
glycoproteins directly in gels. This kit can also be used
to detect glycoproteins after Western transfer to PVDF
membranes.
Rabbit
Mouse
Rabbit
Figure 1. PTM Marker (2 ml of a 6-fold dilution), containing glycosylated and non-glycosylated proteins, was separated by electrophoresis
on a 420% SDS-PAGE gel. The gel was stained for glycoproteins
with GlycoProfile III (left), imaged, and then stained for total protein
with EZBlue Gel Staining Reagent (right). The glycoproteins appear
as bright fluorescent bands. Each band represents approximately
300 ng of protein.
Mouse
Oxidation Reagent
Glycoprotein Staining Reagent
Staining Buffer
sigma-aldrich.com
Specificity
Although this staining procedure is quite selective for
glycoproteins, some non-specific protein staining may
occur and may be more pronounced in some gel formulations. Staining the gel with EZBlue Gel Staining
Reagent after fluorescent imaging will allow identification of non-specifically stained proteins.
An alternative method is to run duplicate gels and fluorescently stain the second gel omitting the oxidation
step. Any fluorescent staining will be non-specific.
17
Glycoprotein Detection
colorimetric TMB or chemiluminescent CPS-1 peroxidase
substrate solutions. Alkaline phosphatase conjugated
streptavidin can also be used to probe the blots. Typical
detection limits for glycoproteins using this is method
are approximately 50-100 ng.
Biotin-hydrazide modification of periodate oxidized glycans can be utilized to label glycoproteins on Western
blots. The blots can then be probed with streptavidinperoxidase.1 Detection can be accomplished with either
Glycan
Periodate
Oxidized
Glycan
O
Periodate
Glycoprotein Detection
Biotin Hydrazide
CH
CH
NH
NH
OH
OH
HN
NH
O=C
C=O
H2C
H2C
CH2
CH2
H2C
H2C
CH2
H
N
S
N
H
CH2
H
N
N
H
Detection Products
B 7639
Biotin Hydrazide
S 5512
S 1878
T 0565
CPS1-60
CPS1-120
CPS1-300
Reference
sigma-aldrich.com
18
Glycoprotein Detection
Glycoprotein Detection
Glycoproteins and
Glycoprotein Standards
There are many methods currently utilized in glycoprotein
detection and identification. Among the most common
are SDS-PAGE, Mass Spectroscopy, HPLC, NMR, and
Western blot. It is helpful and often necessary to include
glycoproteins as standards and/or molecular weight markers.
Invertase is an enzyme that catalyzes the hydrolysis of sucrose into fructose and glucose.
Invertase Glycoprotein Standard is the periplasmic (glycosylated form, external invertase)
with 50% of its mass as polymannan. The periplasmic invertase molecule can exist in a
number of association states each a multiple of the core glycosylated monomer, a 60 kDa
peptide plus oligosaccharide chains and, depending on extraction, purification, and storage
conditions, will exist as a dimer, tetramer, hexamer, or octamer. Since yeast can provide an
alternative system for protein glycosylation that is similar to mammalian systems, periplasmic invertase is often used as a model for the study of the function of oligosaccharides in
glycoproteins and for studies on glycoprotein biosynthesis.
R 1153
8
sigma-aldrich.com
A 5566
Monoclonal Anti-a1-Acid
Glycoprotein antibody
A 0534
A 3418
A 9285
a1-Antichymotrypsin
from human plasma
A 9024
a1-Antitrypsin
from human plasma
Salt-free, lyophilized powder, a1-Proteinase inhibitor, serine protease inhibitor; inhibits trypsin,
chymotrypsin, pancreatic and granulocytic elastase, and acrosin. Effective concentration
equimolar with proteinase. Chromatographically prepared and partially purified.
G 0516
a2-hs-Glycoprotein
from human plasma
minimum 90% (SDS-PAGE) Lyophilized from 20 mM Tris-HCl, pH 8.0, with 200 mM NaCl
A 1960
Aggrecan
from bovine articular cartilage
A 2512
Albumin
from chicken egg white
(Ovalbumin)
A 5503
Albumin
from chicken egg white
(Ovalbumin)
A 5378
Albumin
from chicken egg white
(Ovalbumin)
A 4781
Asialofetuin
from fetal calf serum
19
Glycoprotein Detection
Asialoglycophorin
from human blood Type MN
G 7900
Monoclonal Anti-Glycophorin A
(a) antibody
G 7650
G 7775
Monoclonal Anti-Glycophorin C
(b) antibody
B 9277
R 4011
Produced in mouse, Affinity purified, One unit will bind 1.0 mg of d-biotin
binds to Wrb, a composite blood group antigen resulting from the association between
Glycophorin A and Band 3
A 8706
Avidin
from egg white, recombinant
B 8041
Biglycan
from bovine articular cartilage
C 1063
C 5297
G 4877
Gonadotropin
from pregnant mare serum
C 0755
D 8428
Decorin
from bovine articular cartilage
F 2379
Fetuin
from fetal calf serum
F 3004
Fetuin
from fetal calf serum
Lyophilized powder
Further processing of F 2379 by gel filtration.
F 3879
Fibrinogen
from human plasma
Type I, Contains approx. 15% sodium citrate and approx. 25% sodium chloride.
Approx. 60% protein (80-90% of protein clottable)
F 4385
Fibrinogen
from murine plasma
Contains approx. 20% sodium citrate and approx. 30% sodium chloride.
Approx. 50% protein (over 80% of protein clottable)
F 8630
Fibrinogen
from bovine plasma
Type I-S powder, powder containing approx 75% protein, approx. 10% sodium citrate
and approx. 15% sodium chloride
F 4883
Fibrinogen
from human plasma
Essentially plasminogen-free powder containing approx. 50% protein, approx. 20% sodium
citrate and approx. 30% sodium chloride
H 7017
H 8283
K 3009
sterile-filtered
L 0520
Lactoferrin
from human milk
L 2020
Laminin
from Engelbreth-Holm-Swarm
murine sarcoma
M 1778
Mucin
from porcine stomach
M 3895
Mucin
from bovine submaxillary glands
sigma-aldrich.com
A 9791
Glycoprotein Detection
20
Glycoprotein Detection
sigma-aldrich.com
Glycoprotein Detection
Peroxidase horseradish
Essentially salt-free, Lyophilized powder 250-330 units/mg solid (using pyrogallol) Type VI-A
P 8375
Peroxidase horseradish
P 5661
Plasminogen
from human plasma
e-Aminocaproic acid free, Lyophilized powder containing NaCl, EDTA, lysine, and Tris buffer,
plasmin <0.001 unit/unit plasminogen
T 1001
Thyroglobulin
from bovine thyroid
T 1408
holo-Transferrin bovine
T 4132
holo-Transferrin human
21
sigma-aldrich.com
Chemical Deglycosylation
Strategies
22
Chemical Deglycosylation
Strategies
Key Products
H 2761
Hydrazine
24,451-1
Toluene, Anhydrous
S 6297
Sodium Bicarbonate
A 6404
Acetic Anhydride
C 5893
Cupric Acetate
21,747-6
Dowex 50WX2-400
Peptide
CH2OH
R
O
CH2OH
H
N
O
HO
C
NH
H3C
NH
CH2
O
CH
C
NH2
NH
Hydrazinolysis
C
H3C
Peptide
H
N
O
HO
C
O
sigma-aldrich.com
Re-N-acetylation
CH2OH
R
O
CH2OH
O
HO
NH
H3C
C
O
OH
Regeneration of
reducing
terminus
O
HO
C
NH
NH
H3C
H
N
C
O
CH3
23
Alkaline b-Elimination
Trifluoromethanesulfonic Acid
Release of glycans by alkaline b-elimination utilizes ammonium hydroxide/carbonate or sodium hydroxide (in conjunction with sodium borohydride).
Chemical Deglycosylation
Strategies
References
1. Patel, T.P., and Rarekh, R.B., Meth. Enzymol., 230, 57-66 (1994).
2. Bendiak, B. and Cumming, D.A., Carbohydr. Res., 151, 89 (1986).
3. Makino, Y., et al., J. Biochem., 128, 175-180 (2000).
4. Piller, F., and Piller, V., in Glycobiology: A Practical Approach,
Fukuda, M. and Kobata, A. (Eds), pp. 291-328 (IRL/Oxford Univ.
Press, (Oxford UK 1993)).
5. Burgess, A.J., and Norman, R.I., Eur. J. Biochem. 178, 527-533
(1988).
6. Edge, A.S.B., J. Biochem., 376, 339-350 (2003).
7. Sojar, H.T., and Bahl, O.P., Meth. Enzymol, 138, 41-350 (1987).
8. Edge, A.S.B., et al., Anal. Biochem., 118, 131-137 (1981).
References
1. Carlson, D.M., and Blackwell, C., J. Biol. Chem., 243, 616-626 (1968).
2. Lloyd, K.O., Burchell, J., Kudryashov, V., Yin, B.W.T., and TaylorPapadimitriou, J., J. Biol. Chem., 271, 33325-33334 (1996).
3. Morelle, W., Guyetant, R., and Strecker, G., Carbohydr. Res., 306,
435-443 (1998).
GlycoProfile IV Chemical
Deglycosylation Kit
Product Code PP0510
Visit
sigma-aldrich.com/glycogo
to find out more about the latest
glycobiology products
from Sigma-Aldrich
sigma-aldrich.com
COMING
SOON!
24
Enzymatic Deglycosylation
Strategies
PNGase F
sigma-aldrich.com
25
Enzymatic Deglycosylation
Strategies
High-Mannose Glycans
sigma-aldrich.com
Hybrid Glycans
26
sigma-aldrich.com
Enzymatic Deglycosylation
Strategies
Biantennary Glycans
27
sigma-aldrich.com
Enzymatic Deglycosylation
Strategies
Triantennary Glycans
28
sigma-aldrich.com
Enzymatic Deglycosylation
Strategies
29
Enzymatic Deglycosylation
Strategies
sigma-aldrich.com
30
Enzymatic Deglycosylation
Strategies
In-gel Deglycosylation
Excise protein
of interest
Place gel slice in
siliconized tube
Remove destaining
solution and dry
gel in Speed Vac
Rehydrate with
PNGase F solution
Incubate for
30 min. at 37 C
Add water
Incubate 2 hours to
overnight at 37 C
Remove glycancontaining supernatant.
Analyze if desired.
Glycosylation often leads to problems in subsequent protein analysis procedures. Glycopeptides generally do not
readily ionize during MS analysis leading to insufficient
spectral data. Furthermore, proteolytic digestion of the
native glycoprotein is often incomplete due to steric
hindrance by the oligosaccharides. Removal of the
carbohydrate groups from a glycoprotein prior to
protein identification is preferred.
Sigmas GlycoProfile I Enzymatic In-gel N-Deglycosylation
Kit is optimized to provide a convenient and reproducible
method to N-deglycosylate and tryptically digest protein
samples in 1D or 2D polyacrylamide gel slices for subsequent MS or HPLC analysis. The procedure is suitable for
Coomassie Blue and Colloidal Coomassie stained gels and
may be used with gels silver stained and destained using
Sigmas Proteo Silver Plus Kit (Product Code PROT-SIL2).
GlycoProfile Enzymatic In-gel N-Deglycosylation kit includes
the enzymes and reagents necessary for N-linked deglycosylation and tryptic digestion. The samples can then be
desalted and concentrated for analysis by MALDI-TOF or
electrospray MS.
Features & Benefits
Provides all components for in-gel deglycosylation and
trypsinization of protein samples Conveniently prepares deglycosylated protein samples for analysis by
MS or HPLC
Utilizes PNGase F for the enzymatic removal of N-linked
glycans Proteins remain intact, unlike the use of chemical deglycosylation which can degrade the protein
Includes Proteomics Grade PNGase F and Trypsin
Highly purified enzymes possess no unwanted activities
or additives to complicate analysis
PNGase F is supplied lyophilized from a low salt buffer
Allows reconstitution of the enzyme to any concentration needed
Works in solution or with gel slices Allows choice
of methods
Components
Destaining Solution
Proteomics Grade PNGase F
sigma-aldrich.com
Remove and
discard liquid
31
Enzymatic Deglycosylation
Strategies
In-solution Deglycosylation
Add denaturant
Octyl b-D-Glucopyranoside
Incubate at 37 C for 1 hour
sigma-aldrich.com
2-Mercaptoethanol
32
Enzymatic Deglycosylation
Strategies
CH2OH
R
O
NH
Peptide
CH2OH
R
O
O
HO
PNGase F
NH
CH2
C
NH
H3C
H
N
OH
HO
H3C
Peptide
CH
C
O
NH
CH2
C
O
Peptide
HO
CH
Core GlcNAc
Asparagine Residue
Peptide
PNGase F Products
P 7367
8
PNGase F
sigma-aldrich.com
Proteomics Grade PNGase F from Sigma-Aldrich is extensively purified and contains minimal
residual buffer salts and no glycerol or other stabilizers or preservatives that may interfere
in sensitive glycoprotein analysis methods.
G 5166
8
PNGase F
33
PNGase F
N 3786
a(23,6,8,9) Neuraminidase
PP0200*
8
Glycoprofile I
In-Gel N-Deglycosylation Kit
PP0201*
8
Glycoprofile II
In-Solution N-Deglycosylation Kit
N-DEGLY
The N-DEGLY kit includes Endoglycosidase F1, F2, and F3 as well as an optimized reaction
buffer for each. The kit is intended for use in the deglycosylation of N-linked oligosaccharides
from glycoproteins under native conditions. Particular residues, due to their location in
the native protein structure, are often resistant to the traditional deglycosylation methods
using PNGase F and cannot be removed unless the protein is denatured. Endoglycosidases
F1, F2, and F3 are less sensitive to protein conformation than PNGase F and are more suitable for deglycosylation of native proteins.
Enzymatic Deglycosylation
Strategies
PNGase A
PNGase A (Glycopeptidase A, Peptide N-Glycosidase A),
hydrolyzes oligosaccharides containing a fucose a(1-3)linked to the asparagine-linked N-acetylglucosamine,
commonly found in glycoproteins from plants or parasitic
worms. These types of glycans are resistant to PNGase F.
Like PNGase F the asparagine asparagine residue from
which the glycan is removed is deaminated to aspartic
acid. However, it is ineffective when sialic acid is present
on the N-linked oligosaccharide.
G 0535
PNGase A
G 1163
O-Glycosidase
E-DEGLY
sigma-aldrich.com
Deglycosylation Products
34
Enzymatic Deglycosylation
Strategies
Endoglycosidase F1 cleaves between the two N-acetylglucosamine residues in the N-linked diacetylchitobiose
glycan core of the oligosaccharide, generating a truncated sugar molecule with one N-acetylglucosamine
residue remaining on the asparagine. High mannose
(oligomannose) and hybrid structures can be removed
by Endoglycosidase F1, but not complex, oligosaccharides.
Useful under native or non-denaturing deglycosylation
conditions.
Endoglycosidase F3 cleaves between the two N-acetylglucosamine residues in the N-linked diacetylchitobiose
glycan core of the oligosaccharide, generating a truncated sugar molecule with one N-acetylglucosamine
residue remaining on the asparagine. Endo F3 cleaves
non-fucosylated biantennary and triantennary structures
at a slow rate, but only if peptide-linked. Core fucosylation of biantennary structures increases activity up to
400-fold. Endo F3 has no activity on oligomannose and
hybrid molecules. Endo F3 will also cleave fucosylated
trimannosyl core structures on free and protein-linked
oligosaccharides. Native deglycosylation of complex
tetrantennary glycans requires sequential hydrolysis
down to the trimannosyl-diacetylchitobiose core.
R1 = Asn or H
R2 = Oligomannose or hybrid configurations
R3 = H or a(1-6)fucose
Endoglycosidase F2 cleaves between the two N-acetylglucosamine residues in the N-linked diacetylchitobiose
glycan core of the oligosaccharide, generating a truncated sugar molecule with one N-acetylglucosamine
residue remaining on the asparagine. Endo F2 cleaves
biantennary complex and to a lesser extent high mannose oligosaccharides. Fucosylation has little effect on
Endo F2 cleavage of biantennary structures. Endo F2 will
not cleave hybrid structures. Useful under native or nondenaturing deglycosylation conditions.
R1 = H or Asn
R2 = Biantennary and oligomannose configurations
R3 = H or a(1-6)fucose
sigma-aldrich.com
Endoglycosidase F Products
E 9762
Endoglycosidase F1
E 0639
Endoglycosidase F2
E 2264
Endoglycosidase F3
35
R1
R2
R3
R4
=
=
=
=
Enzymatic Deglycosylation
Strategies
Endoglycosidase H
Oligomannose (2-150)
H or mono- or oligosaccharide at the C2 or C4 position
H or a(1-6)fucose
Asn or Dolichol pyrophosphate
Endoglycosidase H
E 6878
Endoglycosidase H
E 7642
Endoglycosidase H
sigma-aldrich.com
Endoglycosidase H Products
A 0810
36
Glycan Analysis
and Labeling
Glycan Labeling
Chemical derivatization is now the most common method
used for labeling glycans at their reducing ends by reductive amination. A single molecule of fluorescent label can
be incorporated to each mono- or oligosaccharide, thus
allowing molar quantities to be determined. The sensitivity
Technique
HPLC
sigma-aldrich.com
Gel Electrophoresis
Fluorescent Label
Abbreviation
Product Code
2-Aminobenzoic acid
2-AA
10678
2-Aminobenzamide
2-AB
10710
2-Aminopyridine
2-AP
09340
8-Aminonaphthalene-1,3,6-trisulfonic acid
ANTS
08658
2-Aminoacridone
AMAC
06627
Capillary Electrophoresis
9-Aminopyrene-1,3,6-trisulfonic acid
APTS
09341
Mass spectrometry
2-Aminobenzamide
2-AB
10710
37
Fetuin
16,000
12,000
8,000
4,000
0
70
80
90
100
110
120
130
Partially hydrolyzed dextran, as a source of glucose oligomers, is used as an external standard and the elution
position of each peak is expressed in glucose units (gu)
(see Fig. 3). The elution positions of peaks in an unknown
glycan pool are assigned an overall gu value by comparison with the standard dextran ladder. These values may
then be used to predict possible structures for unknown
glycans. For example, it has been reported that the glycans
released from RNase B consist solely of N-linked oligomannose structures. The five peaks obtained upon profiling
of the glycans from RNase B can therefore be assigned
structures ranging from Man-5 to Man-9, as indicated
on Figure 1.
1
4
5
4,400
800
Glycan Analysis
and Labeling
20,000
Fluorescence
RNase B
Fluorescence
Man-5
Fluorescence
600
Man-6
400
3,300
7
2,200
8
9
10
11
1,100
Man-8
200
Man-7
Man-9
0
70
80
90
100
110
120
20
40
60
80
100
120
140
160
sigma-aldrich.com
38
Glycan Analysis
and Labeling
Fetuin glycan
library
Fluorescence
8,000
6,000
Neutral
Di
Tetra
Tri
4,000
Mono
2,000
0
0
10
12
14
16
18
20
22
Neutral glycans
Fluorescence
300
200
100
sigma-aldrich.com
0
60
70
80
90
100
110
120
130
140
C)
400
Monosialylated glycans
300
Fluorescence
200
100
0
60
70
80
90
100
110
120
130
140
39
Glycan Analysis
and Labeling
C
1136.22
100
A
933.20
D
1257.29
H
1745.52
E
1339.30
1152.26
974.16
%
G
1663.48
1298.30
F
1542.41
1419.33
1704.52
1501.41
975.16
J
I
1948.57
1866.59
B
1095.08
990.12
1762.53
991.14
1035.10
1964.66
K
2110.68
L
2151.74
m/z
900
1000
1100
1200
1300
1400
1500
Figure 5. Positive ion MALDI MS of the N-linked glycans from ovalbumin released by treatment with PNGase F. Recorded from 2,5-DHB
1600
1700
1800
1900
2000
2100
2200
2300
0
800
1907.65
1558.40
40
sigma-aldrich.com
Glycan Analysis
and Labeling
41
sigma-aldrich.com
Glycan Analysis
and Labeling
42
Procedure
The following method, commonly referred to as the
dried-droplet method, is based on the original MALDI
experiments and remains the most commonly used
method in the mass spectrometry community.
1. Transfer 3 ml of the appropriate matrix solution
(either DHB or Super DHB at 10 mg/ml) into a 0.5 ml
Eppendorf tube.
2. Add 1 ml of 10 pmol/ml solution of a standard glycan or
an unknown sample solution to the tube containing
the matrix. Vortex to mix.
3. Dispense an aliquot (approximately 1.5 ml) of this
mixture onto the MALDI target plate.
A)
933.32
Glycan Analysis
and Labeling
100
934.32
935.32
936.33
915.16
949.29
937.33
931.11
950.29
m/z
915
920
925
930
935
940
945
950
955
B)
1743.58
100
1744.59
1745.59
%
1746.59
1747.59
1760.56
sigma-aldrich.com
1748.59
0
m/z
1715
1720
1725
1730
1735
1740
1745
1750
1755
1760
1765
1770
1775
43
100
2393.85
2395.85
2396.86
%
2397.86
2398.86
2409.82
m/z
2360
2370
2380
2390
2400
2410
2420
2430
2440
Glycan Analysis
and Labeling
2350
Figure 6. MALDI-TOF mass spectra of (A) Man-3 glycan, (B) Man-8 glycan, and (C) NA4 glycan obtained in positive ion reflectron mode using
DHB as matrix. Each of these glycans are detected as the sodium adduct of the molecular ion [M+Na]+.
COMING
SOON!
Visit
sigma-aldrich.com/glycogo
to find out more about the latest
glycobiology products
from Sigma-Aldrich
Amount Supplied
M 8418
Man-3 Glycan
910.3278
933.3175
1 nmol (approx.)
M 5918
Man-8 Glycan
1720.5919
1743.5817
1 nmol (approx.)
M 8918
NA4 Glycan
2370.8566
2393.8463
1 nmol (approx.)
D 1942
2,5-DHB matrix
2 x 10 mg
D 1817
2 x 10 mg
G 2164
Matrix solvent
(10% ethanol + 10 mM NaCl)
10 ml
sigma-aldrich.com
44
Glycan Analysis
and Labeling
100
831.06
945.43
896.98
1075.39
1299.76
1525.73
1653.00
1997.02
2072.01
2169.84
2313.50
2613.30
0
800
m/z
900 1000 1100 1200 1300 1400 1500 1600 1700 1800 1900 2000 2100 2200 2300 2400 2500 2600 2700 2800 2900
Figure 7. MALDI-TOF mass spectrum of A1 glycan obtained in negative ion linear mode using THAP as matrix and detected as the [M-H] ion.
sigma-aldrich.com
References
1. Stahl, B., Steup, M., Karas, M., and Hillenkamp, F., Anal. Biochem.,
63, 1463-1466 (1991).
2. Karas, M., Ehring, H., Nordhoff, E., Stahl, B., Strupat, K., Hillenkamp,
F., Grehl, M., and Krebs, B., Org. Mass. Spectrom., 28, 1476-1481 (1993).
3. Harvey, D.H., Rapid Commun. Mass Spectrom., 7, 614-619 (1993).
45
Electrophoresis Products
08658
06627
2-Aminoacridone (AMAC)
14,644-7
15,615-9
Sodium cyanoborohydride
D 8779
Dimethylsulfoxide (DMSO)
A 4058
M 7279
N,N-Methylenebisacrylamide (BIS)
T 4904
A 3678
Ammonium sulfate
T 8133
N,N,NN-Tetramethylenediamine (TEMED)
Electrophoresis of Glycans
Glycan Analysis
and Labeling
Glycan Sequencing
Enzymatic analysis of oligosaccharides using highly specific
exoglycosidases, either sequentially or in a matrix array,
is a powerful technique to determine the sequence and
structure of glycan chains. Initially, the glycan pool can
be separated into individual oligosaccharides and each
of these purified glycans can be digested sequentially with
various linkage-specific exoglycosidase enzymes. Figure 9
shows some of the most commonly used exoglycosidase
enzymes used to determine the structures of N-linked
glycans. After appropriate incubation, the product of
digestion can be analyzed by normal phase HPLC or by
MALDI MS. On the HPLC system, structures are assigned to
each peak from the known specificity of the exoglycosidase
used in the digestion and the pre-determined incremental
gu values of individual monosaccharide residues.
Sequencing of oligosaccharides present in a glycan pool
can also be carried out simultaneously on aliquots of the
pool using enzyme arrays. Products of several digestions
are resolved on normal phase HPLC in separate runs and
the shift in the retention times of individual peaks, due
to the action of single enzyme, can be noted. As each
individual exoglycosidase removes additional monosaccharides from the glycan chain, the successive shift in the
retention times, and hence their corresponding gu values,
are obtained. By comparison with the experimentally
determined incremental gu values for the addition of
a particular sugar residue to the standard glycan core
structure, possible structure to each peak present in the
original glycan pool can be assigned. These structural
assignments can further be confirmed by tandem MSn
mass spectrometry.
sigma-aldrich.com
46
Glycan Analysis
and Labeling
Figure 9. Some of the exoglycosidase enzymes commonly used to determine the structure of N-linked glycans.
sigma-aldrich.com
Exoglycosidases
A 6805
b-N-Acetylhexosaminidase
F 8899
a-Fucosidase
F 5884
a-Fucosidase
G 4142
b-Galactosidase
G 0413
b-Galactosidase
M 7257
a-Mannosidase
N 3786
a-2-(3,6,8,9)-Neuraminidase
N 7271
a-2,3-Neuraminidase
47
M 8418
Man-3 Glycan
M 2796
Man-3-F Glycan
M 8912
Man-5 Glycan
Glycan and
Carbohydrate Standards
sigma-aldrich.com
48
M 0420
Man-7 D1 Glycan
M 5918
Man-8 Glycan
sigma-aldrich.com
Glycan and
Carbohydrate Standards
M 4164
49
Man-9 Glycan
M 9919
HYBR glycan
Glycan and
Carbohydrate Standards
N-Linked Hybrid
N-Linked Complex
A1 Glycan
sigma-aldrich.com
M 2425
50
M 5800
A2 Glycan
M 2800
A2F Glycan
sigma-aldrich.com
Glycan and
Carbohydrate Standards
M 3800
51
NA2 Glycan
M 2300
NA2B Glycan
M 9419
NA2F Glycan
sigma-aldrich.com
M 5925
Glycan and
Carbohydrate Standards
N-Linked Complex
52
M 1925
NGA2F Glycan
M 1800
NGA2FB Glycan
sigma-aldrich.com
Glycan and
Carbohydrate Standards
M 1675
53
M 2925
A3 Glycan
M 8793
NA3 Glycan
Glycan and
Carbohydrate Standards
N-Linked Complex
sigma-aldrich.com
54
M 8918
NA4 Glycan
Glycan and
Carbohydrate Standards
M 2175
sigma-aldrich.com
55
N-Asn
A 3432
N2-Asn
A 2292
MN Glycan
A 6587
NM Glycan
A 5065
3a-Fucosyl-N-acetylglucosamine
A 9566
4a-Fucosyl-N-acetylglucosamine
M 1050
2a-Mannobiose
sigma-aldrich.com
A 6681
Glycan and
Carbohydrate Standards
N-Linked Fragments
56
Glycan and
Carbohydrate Standards
N-Linked Fragments
M 6155
4a-Mannobiose
M 7788
6a-Mannobiose
D 5422
3a,6a-Mannotriose
M 0925
3a,6a-Mannopentaose
Syn: a-Man-(13)(a-Man-[16])-a-Man-(16)(a-Man-[13])-Man
112828-69-0, C30H52O26
sigma-aldrich.com
57
N-Acetyllactosamine
A 6919
6b-GlcNAc-D-galactose
A 8297
6b-GlcNAc-lactose
F 0393
2-Fucosyllactose
F 9641
3-Fucosyllactose
L 6770
Lacto-N-tetraose
L 8526
Lacto-N-neo-tetraose
sigma-aldrich.com
A 7791
Glycan and
Carbohydrate Standards
58
sigma-aldrich.com
Glycan and
Carbohydrate Standards
LDFT Glycan
L 5908
Lacto-N-fucopentaose I
L 6401
Lacto-N-fucopentaose II
L 7777
Lacto-N-fucopentaose III
L 7902
Lacto-N-fucopentaose V
59
Lacto-N-difucohexaose I
L 6645
Lacto-N-difucohexaose II
L 6654
Lacto-N-hexaose
L 8283
para-Lacto-N-hexaose
F 5171
Fucosyl-para-lacto-N-hexaose IV
sigma-aldrich.com
L 7033
Glycan and
Carbohydrate Standards
60
sigma-aldrich.com
Glycan and
Carbohydrate Standards
3-Sialyl-3-fucosyllactose
A 4564
LS-Tetrasaccharide a
A 4689
LS-Tetrasaccharide b
A 4814
LS-Tetrasaccharide c
61
D 9045
Disialyllacto-N-tetraose
A 8681
3-Sialyllactose
from bovine colostrum
3-Sialyllactose
from human milk
6-Sialyllactose
from human milk
6-Sialyllactose
from bovine colostrum
A 9079
A 9204
A 8556
Glycan and
Carbohydrate Standards
sigma-aldrich.com
62
Glycan and
Carbohydrate Standards
3-Sialyl-N-acetyllactosamine
A 8431
6-Sialyl-N-acetyllactosamine
A 0828
Sialyllactose
from human milk
A 3307
Sialyllactose
from bovine colostrum
sigma-aldrich.com
63
4a-Galactobiose
G 9662
4b-Galactobiose
A 0167
Galacto-N-biose
A 9150
Lacto-N-biose
F 7012
H-Disaccharide
G 8394
4b-Galactosyllactose
G 9287
Globotriose
sigma-aldrich.com
G 8281
Glycan and
Carbohydrate Standards
64
sigma-aldrich.com
Glycan and
Carbohydrate Standards
A-Trisaccharide
B 1422
B-Trisaccharide
F 7297
H-Trisaccharide
A 8956
A-Tetrasaccharide
Syn: a-D-GalNAc-(13)[a-L-Fuc-(12)]-b-D-Gal-(14)-D-Glc
59957-92-5, C26H45NO20
A 1955
A-Pentasaccharide
Syn: a-D-GalNAc-(13)-[a-L-Fuc-(12)]-b-D-Gal-(14)-[a-L-Fuc-(13)]-D-Glc
50624-46-9, C32H55NO24
65
B 3791
B-Pentasaccharide
Syn: a-Fuc-(12)[a-Gal-(13)]-b-Gal-(14)[a-Fuc-(13)]-Glc
72468-43-0, C30H52O24
A 7560
iso-A-Pentasaccharide
Syn: a-GalNAc-(13)[a-Fuc-(12)]-b-Gal-(13)-[a-Fuc-(14)]-Glc,
A-Leb-Pentasaccharide
128464-25-5, C32H55NO24
B 0799
iso-B-Pentasaccharide
Syn: a-Fuc-(12)[a-Gal-(13)]-b-Gal-(13)[a-Fuc-(14)]-Glc,
b-Leb-Pentasaccharide
128464-26-6, C30H52O24
Glycan and
Carbohydrate Standards
L 7276
Lewis-a trisaccharide
L 5152
Lewis-X trisaccharide
sigma-aldrich.com
66
sigma-aldrich.com
Glycan and
Carbohydrate Standards
Lewis-b tetrasaccharide
L 7784
Lewis-Y tetrasaccharide
L 7401
Lewis-Y hexasaccharide
S 2279
3-Sialyl-Lewis-a tetrasaccharide
S 1782
3-Sialyl-Lewis-X tetrasaccharide
67
3a-Galactobiose
G 9787
3a,4b-Galactotriose
G 9912
3a,4b,3a-Galactotetraose
M 5107
IgM, Lambda
from murine myeloma
M 2521
IgM, Lambda
from murine myeloma
G 7528
D-Glucose
G 6404
D-Galactose
M 8296
D-Mannose
Syn: D-Mannopyranose
3458-28-4, minimum 99% SigmaUltra, C6H12O6
F 2252
L-Fucose
Syn: 6-Deoxy-L-galactose
2438-80-4, minimum 99%, C6H12O5
X 3877
D-Xylose
A 8625
N-Acetyl-D-glucosamine
A 2795
N-Acetyl-D-galactosamine
A 0812
N-Acetylneuraminic acid
Syn: NeuNAc, Sialic acid, Lactaminic acid, NANA, 5-Acetamido-3,5-dideoxy-D-glyceroD-galactononulosonic acid, NAN
131-48-6, Type IV-S, C11H19NO9
G 2755
N-Glycolylneuraminic acid
Syn: NeuNGl
1113-83-3, C11H19NO10
G 8645
Glycan and
Carbohydrate Standards
Monosaccharides
sigma-aldrich.com
68
sigma-aldrich.com/glycogo
Glycobiology
Product Directory
sigma-aldrich.com
For advice and information on our products, contact Sigma Technical Services by email at
techserv@sial.com
69
Enzymes
Complete List of Glycolytic Enzymes
A 9763
a-N-Acetylgalactosaminidase
A 6680
Expressed in E. coli
A 6306
Agarase
A 4551
a-Amylase
A 6814
a-Amylase
A 6380
a-Amylase
A 6211
a-Amylase
A 2771
a-Amylase
A 0521
a-Amylase
A 1031
a-Amylase
A 4268
a-Amylase
A 6255
a-Amylase
A 2643
a-Amylase
A 3403
a-Amylase
A 3306
a-Amylase
Heat-stable, solution
A 7130
b-Amylase
A 7005
b-Amylase
I 2758
Isoamylase
Amylase
A 7420
Amyloglucosidase
A 7095
Amyloglucosidase
A 2330
Amyloglucosidase
A 7255
Amyloglucosidase
A 3042
Amyloglucosidase
A 3514
Amyloglucosidase
C 1184
Cellulase
C 2605
Cellulase
C 2730
Cellulase
C 9422
Cellulase
C 0898
Cellulase
C 7809
Chitinase
C 1650
Chitinase
C 6137
Chitinase
C 1525
Chitinase
C 9830
Chitosanase
C 0794
Chitosanase
Glycobiology
Product Directory
Amyloglucosidase
Cellulase
Chitinase
Chondroitinase
C 2905
Chondroitinase ABC
C 3667
Chondroitinase ABC
sigma-aldrich.com
Chitosanase
70
Enzymes
Complete List of Glycolytic Enzymes
Chondroitinase (cont)
C 2262
Chondroitinase AC
C 2780
Chondroitinase AC
C 0954
Chondroitinase C
C 8058
Chondroitinase B
Chondrosulfatase
C 2655
Chondro-4-sulfatase
C 3030
Chondro-6-sulfatase
C 8274
Cytohelicase
D 9909
Dextran sucrase
D 4668
Dextranase
D 5884
Dextranase
D 8144
Dextranase
D 9515
Driselase
From Basidiomycetes sp., crude powder containing laminarinase, xylanase and cellulase
Dextranase
Glycobiology
Product Directory
Endoglycosidase
E 9762
Endoglycosidase F1
E 0639
Endoglycosidase F2
E 2264
Endoglycosidase F3
E 6878
Endoglycosidase H
A 0810
Endoglycosidase H
E 7642
Endoglycosidase H
F 1924
a1-(2,3,4)-Fucosidase
F 9272
a1,2-Fucosidase
F 3023
a1-(3,4)-Fucosidase
F 6272
a1,6-Fucosidase
F 8899
a-Fucosidase
F 6151
a-L-Fucosidase
F 5884
a-L-Fucosidase
Fucosidase
sigma-aldrich.com
Galactosidase
G 7163
a-Galactosidase
G 4408
a-Galactosidase
G 8507
a-Galactosidase
G 1288
b1-(3,4,6)-Galactosidase
G 0288
b1-(3,6)-Galactosidase
G 0413
b1,4-Galactosidase
G 0914
b1,6-Galactosidase
G 3153
b-Galactosidase
G 5635
b-Galactosidase
G 5160
b-Galactosidase
G 4142
b-Galactosidase
G 4155
b-Galactosidase
71
Enzymes
Complete List of Glycolytic Enzymes
Galactosidase (cont)
G 6008
b-Galactosidase
G 2513
b-Galactosidase
G 1875
b-Galactosidase
From bovine liver, grade III, lyophilized powder, approx. 0.15 units/mg
G 3782
b-Galactosidase
G 5507
Galactosyltransferase
G 2133
Glucose Oxidase
G 8162
b-Glucuronidase
G 7396
b-Glucuronidase
G 7646
b-Glucuronidase
G 7771
b-Glucuronidase
G 7896
b-Glucuronidase
G 5897
b-Glucuronidase
G 8271
b-Glucuronidase
G 0251
b-Glucuronidase
G 0501
b-Glucuronidase
G 0376
b-Glucuronidase
G 0751
b-Glucuronidase
G 0876
b-Glucuronidase
G 8885
b-Glucuronidase
G 0762
b-Glucuronidase
From Helix pomatia, type H-3AF, crude solution, minimum 60,000 units/ml
G 1512
b-Glucuronidase
G 7017
b-Glucuronidase
G 7770
b-Glucuronidase
G 8132
b-Glucuronidase
From Patella vulgata (keyhole limpet), type L-II, lyophilized powder, 1,000,000-3,000,000 units/g
G 4259
b-Glucuronidase
Glycobiology
Product Directory
Glucuronidase
Glucosaminidase
A 7708
b-N-Acetylglucosaminidase
A 2415
b-N-Acetylglucosaminidase
A 6152
b-N-Acetylglucosaminidase
A 6805
b-N-Acetylglucosaminidase
A 2264
b-N-Acetylglucosaminidase
A 3189
b-N-Acetylglucosaminidase
A 3391
b-N-Acetylglucosaminidase A
A 7640
b-N-Acetylglucosaminidase B
G 3651
a-Glucosidase
G 0660
a-Glucosidase
G 5003
a-Glucosidase
G 6906
b-Glucosidase
G 4511
b-Glucosidase
G 0395
b-Glucosidase
sigma-aldrich.com
Glucosidase
72
Enzymes
Complete List of Glycolytic Enzymes
G 9028
d4,5-Glycuronidase
G 1163
O-Glycosidase
H 2125
Hemicellulase
H 2519
Heparinase I
H 6512
Heparinase II
H 8891
Heparinase III
Heparinase
Hesperidinase
H 8137
Hesperidinase
H 8510
Hesperidinase
Glycobiology
Product Directory
Hyaluronidase
H 3506
Hyaluronidase
H 3884
Hyaluronidase
H 3757
Hyaluronidase
From bovine testes, type VIII, lyophilized powder, approx. 300 units/mg
H 3631
Hyaluronidase
H 2126
Hyaluronidase
H 2251
Hyaluronidase
H 6254
Hyaluronidase
H 1136
Hyaluronidase
I 9274
Invertase
I 9253
Invertase
I 4504
Invertase
From bakers yeast (S. cerevisiae), grade VII, minimum 400 units/mg
I 4753
Invertase
L 9903
Lacto-N-biosidase
Invertase
Laminarinase
L 9259
Laminarinase
L 5272
Laminarinase
L 7773
Lysozyme
L 7001
Lysozyme
L 6876
Lysozyme
L 6394
Lysozyme
L 8402
Lysozyme
L 1129
Lysozyme Agarose
M 1266
Lysozyme Agarose
L 2879
From chicken egg white, grade VI, not dialyzed or lyophilized, approx. 60,000 units/mg
sigma-aldrich.com
Lysozyme
Mannosidase
M 7257
a-Mannosidase
M 2658
a-Mannosidase
M 9400
b1,4-Mannosidase, b-Mannosidase
From Helix pomatia, From snail acetone powder, ammonium sulfate suspension, 5-30 units/ml
From Canavalia ensiformis (jack bean), ammonium sulfate suspension, approx. 20 units/mg
73
Enzymes
Complete List of Glycolytic Enzymes
N 1385
Naringinase
N 7271
a2,3-Neuraminidase
N 5521
a2-(3,6)-Neuraminidase
N 8271
a2-(3,6,8,9)-Neuraminidase
N 3786
a2-(3,6,8,9)-Neuraminidase
Proteomics grade
N 4883
Neuraminidase Agarose
From Clostridium perfringens (C. welchii), type X-A, ammonium sulfate suspension,
20-30 units/g agarose
N 5254
Neuraminidase Agarose
From Clostridium perfringens (C. welchii), type VI-A, ammonium sulfate suspension
N 2133
Neuraminidase
N 2876
Neuraminidase
From Clostridium perfringens, 0.5-6 units/mg solid, From Vibrio cholerae, type III,
1-3 units/mg protein
N 5631
Neuraminidase
From Clostridium perfringens (C. welchii), type VIII, lyophilized powder, 10-20 units/mg
N 3001
Neuraminidase
N 7771
Neuraminidase
N 3642
Neuraminidase
N 7885
Neuraminidase
N 6514
Neuraminidase
C 6105
Novozyme 188
P 7052
P 2611
Pectinase
P 2736
Pectinase
P 2401
Pectinase
P 0764
Pectinesterase
P 3026
Pectolyase
P 5431
Pectolyase
P 8375
Peroxidase, Horseradish
Essentially salt-free
G 5166
PNGase F
P 3304
Polygalacturonase
P 5420
Pullulanase
P 1067
Pullulanase
S 9754
Sulfatase
S 1629
Sulfatase
S 9626
Sulfatase
S 8504
Sulfatase
From Patella vulgata (keyhole limpet), type IV, essentially salt-free, lyophilized powder,
10 units/mg
Glycobiology
Product Directory
Neuraminidase
Pectinase
Pectolyase
Pullulanase
sigma-aldrich.com
Sulfatase
74
Enzymes
Complete List of Glycolytic Enzymes
Sulfatase (cont)
S 8629
Sulfatase
From Patella vulgata (keyhole limpet), type V, Essentially salt-free, lyophilized powder,
minimum 5 units/mg
S 1062
2-O-Sulfatase
S 9751
Sulfatase
From Helix pomatia, type H-2, crude solution, minimum 2,000 units/ml
T 4528
Thioglucosidase
T 8778
Trehalase
From porcine kidney, buffered aqueous glycerol solution, minimum 0.4 units/mg
V 2010
Viscozyme L
X 4001
Xylanase
X 2753
Xylanase
X 3876
Xylanase
X 1378
b1,4-Xylosidase
X 3501
b-Xylosidase
Xylanase
Xylosidase
Glycobiology
Product Directory
Ceramide Glycanase
E 2277
Endoglycoceramidase II
plus activator II
From Rhodococcus sp., endo-type cleavage of cell surface glycosphingolipids of living cells
without damaging other cell components
P 5542
Phospholipase C,
Phosphatidylinositol-specific
P 8804
Phospholipase C,
Phosphatidylinositol-specific
P 5147
Pronase E
sigma-aldrich.com
Glycosyltransferases
74188
a-1,3-Galactosyl-Transferase Kit
77038
a-1,3-Galactosyl-Transferase
59505
b-1,4-Galactosylation Kit
90261
b-1,4-Galactosyl Transferase I
48279
Galactosyltransferase
48281
Galactosyltransferase
75
Inhibitors
Inhibitors of Carbohydrate Metabolism
P 7340
()-threo-1-Phenyl-2-decanoyl-amino-3-morpholino-1-propanol hydrochloride
Syn: PDMP
H 9632
(2R,5R)-Bis(hydroxymethyl)-(3R,4R)-dihydroxypyrrolidine
Syn: DMDP
D 8390
1,4-Dideoxy-1,4-imino-D-mannitol hydrochloride
D 9160
1-Deoxymannojirimycin hydrochloride
D 9305
1-Deoxynojirimycin hydrochloride
A 5416
2-Acetamido-1,2,5-trideoxy-1,5-imino-D-glucitol
D 8875
Deoxyglucose
D 8375
2-Deoxy-D-glucose
D 6134
2-Deoxy-D-glucose
D 3179
2-Deoxy-D-glucose
SigmaUltra
B 1147
6-Bromo-4-cyclohexene-1,2,3-triol
Mixed isomers
B 6676
7(S)-Brefeldin A-7-3H
Ethanol solution
B 4894
Benzyl 2-acetamido-2-deoxy-a-D-galactopyranoside
Syn: a-D-GalNAc-1OCH2Ph
B 6542
Brefeldin A
B 7651
Brefeldin A
C 3784
Castanospermine
27689
Concanamycin A
C 9705
Concanamycin A
C 5424
Conduritol B epoxide
Syn: 1,2-Anhydro-myo-inositol
D 9641
Deoxygalactonojirimycin hydrochloride
Syn: 1,5-Dideoxy-1,5-imino-D-galactitol
P 4194
DL-threo-1-Phenyl-2-palmitoyl-amino-3-morpholino-1-propanol
69895
M 5273
Syn: Monensin A
D 9050
N-Acetyl-2,3-dehydro-2-deoxyneuraminic acid
N 8028
Nikkomycin Z
B 8299
N-Butyldeoxynojirimycin
M 1777
N-Methyl-1-deoxynojirimycin
S 6640
Swainsonine
S 9263
Swainsonine
Synthetic
T 0527
Tunicamycin A1 homolog
Approx. 95%
T 1152
Tunicamycin C2 homolog
T 7765
Tunicamycin
Brefeldin
Glycobiology
Product Directory
Con A
Monensin
Nojirimycin
Swainsonine
sigma-aldrich.com
Tunicamycin
76
Substrates
Glycolytic Enzyme Substrates
Acetylneuraminic
M 8639
2-(4-Methylumbelliferyl)-a-D-N-acetylneuraminic acid
N 1266
2-O-(o-Nitrophenyl)-a-D-N-acetylneuraminic acid
Approx. 95%
N 1516
2-O-(p-Nitrophenyl)-a-D-N-acetylneuraminic acid
Approx. 95%
B 4666
Arabinofuranoside
N 3641
4-Nitrophenyl a-L-arabinofuranoside
N 3512
4-Nitrophenyl a-L-arabinopyranoside
N 0520
4-Nitrophenyl b-L-arabinopyranoside
Cellobioside
N 4764
2-Nitrophenyl b-D-cellobioside
M 6018
4-Methylumbelliferyl b-D-cellobioside
N 5759
4-Nitrophenyl b-D-cellobioside
B 6795
5-Bromo-4-chloro-3-indolyl b-D-cellobioside
Syn: X-Cellobiose
N 0145
4-Nitrophenyl b-D-cellopentaoside
N 1262
4-Nitrophenyl b-D-cellotetraoside
Minimum 75%
N 4889
4-Nitrophenyl b-D-cellotrioside
Approx. 95%
M 9763
4-Methylumbelliferyl b-D-N,N-diacetylchitobioside
N 6133
4-Nitrophenyl N,N-diacetyl-b-D-chitobioside
N 8638
4-Nitrophenyl b-D-N,N,N-triacetylchitotriose
M 5639
4-Methylumbelliferyl b-D-N,N,N-triacetylchitotrioside
Chitobioside
Glycobiology
Product Directory
Fucopyranoside
N 3628
4-Nitrophenyl a-L-fucopyranoside
N 3378
4-Nitrophenyl b-D-fucopyranoside
N 2505
4-Nitrophenyl b-L-fucopyranoside
B 9511
5-Bromo-4-chloro-3-indolyl b-D-fucopyranoside
Minimum 98%
B 2280
5-Bromo-4-chloro-3-indolyl b-L-fucopyranoside
Minimum 98%
N 3253
2-Nitrophenyl b-D-fucopyranoside
M 5510
4-Methylumbelliferyl b-D-fucoside
Minimum 98%
M 4508
4-Methylumbelliferyl b-L-fucoside
Minimum 98%
Fucoside
sigma-aldrich.com
Galactopyranoside
67610
1-Methyl-3-indolyl-b-D-galactopyranoside
N 7508
1-Naphthyl a-D-galactopyranoside
N 2259
2-Naphthyl b-D-galactopyranoside
N 2509
2-Nitrophenyl 1-thio-b-D-galactopyranoside
N 8888
2-Nitrophenyl a-D-galactopyranoside
N 1127
2-Nitrophenyl b-D-galactopyranoside
N 6129
3-Nitrophenyl a-D-galactopyranoside
Minimum 98%
C 7553
4-Chloro-3-indolyl b-D-galactopyranoside
M 7633
4-Methylumbelliferyl a-D-galactopyranoside
M 1633
4-Methylumbelliferyl-b-D-galactopyranoside
N 2257
4-Nitrophenyl 1-thio-b-D-galactopyranoside
Minimum 95%
N 2766
4-Nitrophenyl 2-acetamido-2-deoxy-3-O-(2-acetamido-2-deoxyb-D-glucopyranosyl)-a-D-galactopyranoside
Minimum 98%
N 3016
4-Nitrophenyl 2-acetamido-2-deoxy-3-O-(b-D-galactopyranosyl)a-D-galactopyranoside
Minimum 98%
N 7643
4-Nitrophenyl 6-O-b-D-galactopyranosyl-b-D-galactopyranoside
Minimum 95%
N 0877
4-Nitrophenyl a-D-galactopyranoside
77
Substrates
Glycolytic Enzyme Substrates
Galactopyranoside (cont)
N 1252
4-Nitrophenyl b-D-galactopyranoside
Minimum 98%
T 3661
4-Trifluoromethylumbelliferyl b-D-galactopyranoside
B 2904
5-Bromo-3-indolyl b-D-galactopyranoside
B 4387
5-Bromo-3-indolyl b-D-galactopyranoside
Minimum 98%
B 6024
5-Bromo-4-chloro-3-indolyl b-D-galactopyranoside
5 mg tablet
B 9146
5-Bromo-4-chloro-3-indolyl b-D-galactopyranoside
Minimum 98%
B 4252
5-Bromo-4-chloro-3-indolyl b-D-galactopyranoside
16669
5-Bromo-6-chloro-3-indolyl-b-D-galactopyranoside
57897
5-Iodo-3-indolyl-b-D-galactopyranoside
36931
6,8-Difluoro-4-methylumbelliferyl-b-D-galactopyranoside
24904
6-Chloro-3-indolyl-b-D-galactopyranoside
F 2756
Fluorescein di-(b-D-galactopyranoside)
Minimum 98%
F 6542
Fluorescein mono-b-D-galactopyranoside
Approx. 90%
I 7513
Indoxyl b-D-galactopyranoside
Minimum 98%
N 2885
N 0257
2-Nitrophenyl N-acetyl-a-D-galactosaminide
N 3273
2-Nitrophenyl-N-acetyl-b-D-galactosaminide
M 9659
4-Methylumbelliferyl N-acetyl-b-D-galactosaminide
N 4264
4-Nitrophenyl N-acetyl-a-D-galactosaminide
Minimum 98%
N 9003
4-Nitrophenyl N-acetyl-b-D-galactosaminide
Minimum 98%
B 3166
5-Bromo-4-chloro-3-indolyl N-acetyl-b-D-galactosaminide
Approx. 95%
Glycobiology
Product Directory
Galactosaminide
Galacturonide
N 8755
4-Nitrophenyl b-D-galacturonide
Glucopyranoside
N 2007
2-Naphthyl b-D-glucopyranoside
N 8016
2-Nitrophenyl b-D-glucopyranoside
M 9766
4-Methylumbelliferyl a-D-glucopyranoside
M 3633
4-Methylumbelliferyl b-D-glucopyranoside
M 0662
4-Methylumbelliferyl-7-(6-sulfo-2-acetamido-2-deoxyb-D-glucopyranoside)
N 2381
4-Nitrophenyl 1-thio-b-D-glucopyranoside
Approx. 95%
N 5513
4-Nitrophenyl 2-acetamido-2-deoxy-3-O-b-D-galactopyranosylb-D-glucopyranoside
N 1377
4-Nitrophenyl a-D-glucopyranoside
Minimum 99%
N 7006
4-Nitrophenyl b-D-glucopyranoside
B 4527
5-Bromo-4-chloro-3-indolyl b-D-glucopyranoside
Approx. 98%
36937
6,8-Difluoro-4-methylumbelliferyl-b-D-glucopyranoside
F 4521
Fluorescein di-(b-D-glucopyranoside)
Minimum 90%
M 9881
4-Methylumbelliferyl N-acetyl-a-D-glucosaminide
M 2133
4-Methylumbelliferyl N-acetyl-b-D-glucosaminide
N 8759
4-Nitrophenyl N-acetyl-a-D-glucosaminide
Minimum 98%
N 9376
4-Nitrophenyl N-acetyl-b-D-glucosaminide
98-100%
B 3041
5-Bromo-4-chloro-3-indolyl N-acetyl-b-D-glucosaminide
Minimum 98%
N 4006
I 6893
Indoxyl b-D-glucoside
I 3750
Indoxyl b-D-glucoside
Minimum 97%
Glucoside
sigma-aldrich.com
Glucosaminide
78
Substrates
Glycolytic Enzyme Substrates
Glucuronide
N 9013
1-Naphthyl b-D-glucuronide
N 2645
2-Nitrophenyl b-D-glucuronide
M 9130
4-Methylumbelliferyl b-D-glucuronide
M 5664
4-Methylumbelliferyl b-D-glucuronide
SigmaUltra, hydrate
N 1627
4-Nitrophenyl b-D-glucuronide
T 6410
4-Trifluoromethylumbelliferyl glucuronide
B 8049
5-Bromo-4-chloro-3-indolyl b-D-glucuronide
B 0522
5-Bromo-4-chloro-3-indolyl b-D-glucuronide
B 3783
5-Bromo-4-chloro-3-indolyl b-D-glucuronide
B 6650
5-Bromo-4-chloro-3-indolyl b-D-glucuronide
B 8174
5-Bromo-4-chloro-3-indolyl b-D-glucuronide
B 5285
5-Bromo-4-chloro-3-indolyl b-D-glucuronide
B 4782
5-Bromo-4-chloro-3-indolyl b-D-glucuronide
B 4532
5-Bromo-6-chloro-3-indolyl b-D-glucuronide
B 4657
5-Bromo-6-chloro-3-indolyl b-D-glucuronide
36939
6,8-Difluoro-4-methylumbelliferyl-b-D-glucuronide
24907
6-Chloro-3-indolyl-b-D-glucuronide
I 7638
Indoxyl b-D-glucuronide
Cyclohexylammonium salt
N 1875
P 0376
Phenolphthalein b-D-glucuronide
Sodium salt
P 0501
Phenolphthalein b-D-glucuronide
Free acid
Glycobiology
Product Directory
Lactopyranoside
M 2405
4-Methylumbelliferyl b-D-lactopyranoside
N 1752
4-Nitrophenyl b-D-lactopyranoside
Maltopentaoside
N 1519
4-Nitrophenyl-a-D-maltopentaoside
Minimum 98%
N 5885
4-Nitrophenyl a-D-maltoside
N 1884
4-Nitrophenyl b-D-maltoside
Maltoside
Mannopyranoside
M 3657
4-Methylumbelliferyl a-D-mannopyranoside
M 0905
4-Methylumbelliferyl b-D-mannopyranoside
N 2127
4-Nitrophenyl a-D-mannopyranoside
N 1268
4-Nitrophenyl b-D-mannopyranoside
Approx. 98%
B 4526
5-Bromo-4-chloro-3-indolyl a-D-mannopyranoside
Approx. 95%
G 2000
2-(b-D-Galactosidoxy)naphthol AS-LC
A 8750
D-()-Salicin
Naphthol
Salicin
S 0625
Rhamnopyranoside
sigma-aldrich.com
N 7763
4-Nitrophenyl a-L-rhamnopyranoside
Xylopyranoside
N 3629
2-Nitrophenyl b-D-xylopyranoside
M 7008
4-Methylumbelliferyl-b-D-xylopyranoside
N 1895
4-Nitrophenyl a-D-xylopyranoside
N 2132
4-Nitrophenyl b-D-xylopyranoside
79
Neoglycoproteins
Neoglycoproteins
Cellulose
A 8585
Cellobiose-BSA
Galactose
A 5908
2-Amido-GalN-BSA
A 2420
a-Gal-FITC-BSA
A 6544
a-Gal-TRITC-BSA
A 8165
b-Gal-FITC-BSA
A 1159
Aminophenyl-b-GalNAc-BSA
Glucose
A 6158
GlcNAc-BSA
A 5543
a-Glc-FITC-BSA
A 1034
Aminophenyl-b-GlcNAc-BSA
A 7172
b-Glc-FITC-BSA
A 6052
GlcNAc-resorufin-BSA
Fructose
A 8426
Fructosamine-BSA (glycated)
A 8301
Fructosamine-HSA (glycated)
A 4042
1-Amido-Fuc-biotin-BSA
A 6033
1-Amido-Fuc-BSA
A 5793
a-L-Fuc-FITC-BSA
A 5918
a-L-Fuc-TRITC-BSA
Glycobiology
Product Directory
Fucose
Lactose
A 8210
Lactose-BSA
A 7799
b-Lactose-biotin-BSA
A 8040
b-Lactose-FITC-BSA
A 7665
b-Lactose-TRITC-BSA
A 5783
Lactitol-1-N-BSA
Maltose
A 5283
Maltitol-1-N-BSA
A 8460
Maltose-BSA
Mannose
A 7924
a-Man-biotin-BSA
A 8303
a-Man-BSA
A 7790
a-Man-FITC-BSA
A 7915
a-Man-TRITC-BSA
A 4664
Aminophenyl-a-Man-BSA
A 3955
b-Xyl-FITC-BSA
sigma-aldrich.com
Xylose
80
Lectins
Lectins are proteins or glycoproteins from non-immune origins that agglutinate cells and/or precipitate complex carbohydrates. The agglutination activity of these highly specific carbohydrate-binding molecules is usually inhibited by a
simple monosaccharide, but for some lectins di-, tri-, and even polysaccharides are required.
Sigma offers a wide range of lectins suitable for the following applications:
Carbohydrate studies
Lymphocyte subpopulation studies
Blood group typing
Fractionation of cells and other particles
Mitogenic stimulation
Histochemical studies
sigma-aldrich.com
Glycobiology
Product Directory
Lectins are isolated from a wide variety of natural sources. Sigma offers a range of lectins suitable for all of the
applications noted above.
Common Name
Taxonomic Name
Common Name
Taxonomic Name
Abrin
Abrus precatorius
Labumum, Scotch
Labumum polyphemus
Asparagus pea
Tetragonolobus purpureas
Lentil
Lens culinaris
Avocado
Perseau americana
Lotus
Tetragonolobus purpureas
Momordica charantia
Mistletoe, European
Viscum album
Broad bean
Vicia faba
Mung bean
Vigna radiata
Bauhinia purpurea
Mushroom
Agaricus bisporus
Castor bean
Ricinus communis
Osage orange
Maclura pomifera
Chick pea
Cicer arietinum
Pagoda tree
Sophora japonica
Cobra, Mozambique
Pea, garden
Pisum sativum
Cobra, Thailan
Peanut
Arachis hypogaea
Con A
Canavalia ensiformis
Pokeweed
Phytolacca americana
Concanavalin A
Canavalia ensiformis
Potato
Solanum tuberosum
Erythrina corallodendron
Phaseolus vulgaris
Daffodil
Narcissus pseudonarcissus
Ptilota plumosa
Eel
Anguilla anguilla
Roman snail
Helix pomatia
Elder
Sambucus nigra
Phaseolus coccineus
Fava bean
Vicia faba
Caragana arborescens
Furze
Ulex europaeus
Snail, garden
Helix aspersa
Gorse
Ulex europaeus
Snowdrop
Galanthus nivalis
Codium fragile
Soybean
Glycine max
Hairy vetch
Vicia villosa
Spindle tree
Euonymus europaeus
Horse gram
Dolichos biflorus
Sweet pea
Lathyrus odoratus
Horseshoe crab
Limulus polyphemus
Thorn Apple
Datura stramonium
Jacalin
Artocarpus intefrifolia
Tomato
Lycopersicon esculentum
Jack bean
Canavalia ensiformis
Wheat germ
Triticum vulgaris
Japanese wisteris
Wisteria floribanda
Winged bean
Psophocarpus tetragonolobus
Jequity bean
Abrus precatorius
Winged pea
Tetragonolobus purpureas
Jimson weed
Datura stramonium
Many lectins are available as conjugates. Conjugation does not alter the specificity of the lectin. The degree of conjugation
is expressed as 1) a mole of dye per mole of lectin basis 2) a unit of enzyme activity per mg of protein or 3) a mg of
protein per ml of packed gel basis. For colloidal gold conjugates, the activity of the lectin is determined by a dot blot
assay using an appropriate glycoprotein. Agglutination activity is expressed in microgram per ml and is determined
from serial dilution studies of a 1 mg per ml solution. This activity is the lowest concentration of lectin to agglutinate
a 2% suspension of appropriate erythrocytes after 1 hour incubation at 25 C.
81
Lectins
Lectins
Product
Number
Source
Subunits
Specificity
Blood Group
Specificity Sugar
L 5640
Agaricus bisporus
58.5
b-Gal(13)GalNAc
L 0881
Arachis hypogaea
120
b-Gal(13)GalNAc
42
a-GalOMe
114
A,B
a-Gal, a-GalNAc
114
a-Gal
L 7759
Peroxidase
L 6135
Biotin
L 7381
FITC
L 3766
L 3515
Mitogenic
Activity
TRITC
Artocarpus integrifolia
L 4650
Peroxidase
L 5147
Agarose
(+)
BS-I
L 3759
Biotin
L 9381
FITC
L 5264
TRITC
L 0890
FITC
L 3019
BS-I-B4
L 5391
Peroxidase
L 2140
Biotin
L 2895
FITC
L 9637
Caragana arborescens
60;120(c)
2;4
GalNAc
L 3141
Cicer arietinum
44
fetuin
L 2638
Codium fragile
60
GalNAc
C 7275
Concanavalin A
102
a-Man, a-Glc
(+)
L 6397
Peroxidase
C 2272
Biotin
C 7642
FITC
C 6904
Agarose
C 9017
Sepharose
L 5021
Gold, 10 nm
L 3642
Gold, 20 nm
C 7898
Ferritin
51
a-Man, a-Glc
(+) (d)
L 9385
Succinyl-Concanavalin A
FITC
L 2766
Datura stramonium
86
(GlcNAc)2
L 2785
Dolichos biflorus
140
A1
a-GalNAc
L 1287
Peroxidase
L 6533
Biotin
L 9142
FITC
L 9658
TRITC
L 9894
Agarose
L 5390
Erythrina cristagalli
56.8
b-Gal(14)GlcNAc
L 7400
Euonymus europaeus
166
B,H
a-Gal(13)Gal
L 8725
Galanthus nivalis
52
(h)
non-reduc. D-Man
L 8775
Agarose
(+)
sigma-aldrich.com
L 3885
Glycobiology
Product Directory
Bandeiraea simplicifolia
L 2380
82
Lectins
Lectins
Product
Number
Source
Subunits
Specificity
Blood Group
Specificity Sugar
Mitogenic
Activity
L 1395
Glycine max
110
GalNAc
(+) (b)
L 2650
Peroxidase
L 4511
TRITC
79
GalNAc
79
GalNAc
49
a-Man
18
NeuNAc
L 6635
L 8764
Biotin
L 3382
Helix pomatia
L 6387
Peroxidase
L 6512
Biotin
L 1034
FITC
L 1261
L 9267
TRITC
Lens culinaris
L 4143
Biotin
L 9262
FITC
L 0511
Limulus polyphemus
400
L 2886
Lycopersicon esculentum
71
L 0401
L 8025
L 4401
L 3138
L 4389
(+)
Agarose
L 2263
L 0651
Glycobiology
Product Directory
Helix aspersa
(GlcNAc)3
(+) (e)
sialic acid
(+)
Biotin
FITC
Maackia amurensis
130
2(a and b)
112
128
oligosaccharide
(+)
126
oligosaccharide
(+)
(GlcNAc)3
(+)
a-Man
(+)
Peroxidase
Phaseolus coccineus
TRITC
Phaseolus vulgaris
L 8629
L 6139
PHA-E
TRITC
L 2769
PHA-L
L 8754
PHA-P
L 2646
PHA-M
L 9379
Phytolacca americana
32(f)
L 5380
Pisum sativum
49
L 0770
35
GalNAc, Gal
120
b-Gal
FITC
L 9895
L 2138
Psophocarpus tetragonolobus
L 3139
Peroxidase
L 3014
Biotin
L 3264
Gal
FITC
Ricinus communis
sigma-aldrich.com
L 7886
Agglutinin, RCA120
L 2758
Peroxidase
L 9514
Ricin, A chain
L 4022
L 9639
Ricin, B chain
L 6890
Sambucus nigra
L 4266
Solanum tuberosum
83
Lectins
Lectins
Product
Number
Source
Subunits
Specificity
Blood Group
Specificity Sugar
L 9254
Tetragonolobus purpureas
120(A), 58(B),
117(C)
4;2;4
a-L-fuc
36
L 5759
Peroxidase
L 3134
Biotin
L 5644
FITC
L 9640
Triticum vulgaris
L 3892
Peroxidase
L 0390
Peroxidase (inactive)
L 5142
Biotin
L 4895
FITC
L 9884
Evans Blue
L 1894
Gold, 10 nm
L 1882
Agarose
(GlcNAc)2, NeuNAc
Mitogenic
Activity
(+)
L 8146
Peroxidase
L 8262
Biotin
L 9006
FITC
L 4889
TRITC
L 4011
Vicia villosa
L 9388
Agarose
L 7513
Isolectin B4
L 7888
Agarose
L 2662
68
a-L-Fuc
139
4(a)
A1+Tn
GalNAc
143
Tn
GalNAc
Viscum album
115 g
b-Gal
Wisteria floribunda
68
GalNAc
34
GalNAc
L 1516
Biotin
L 2016
Reduced
L 1766
Biotin
Notes:
a Subunits are of different molecular weights
b Mitogenic for neuraminidase-treated lymphocytes
c Concentration-dependent mol. wt. change
d Non-agglutinating and mitogenic
sigma-aldrich.com
UEA I
Glycobiology
Product Directory
Ulex europaeus
L 5505
88
sigma-aldrich.com
Obsession with
SERVICE
Passion for
PROCESS
IMPROVEMENT
Come to Sigma-Aldrich
for Answers to Your Questions
Visit us on line at sigma-aldrich.com,
our award-winning Web site
featuring new products, over
2 million Certificates of Analysis
and over 90,000 MSDSs. You can
also search by product name,
CAS number, chemical structures and substructures.
Leadership in
Argentina
SIGMA-ALDRICH DE ARGENTINA, S.A.
Tel: 54 11 4556 1472
Fax: 54 11 4552 1698
France
SIGMA-ALDRICH CHIMIE S..r.l.
Tel appel gratuit: 0800 211 408
Fax appel gratuit: 0800 031 052
Japan
SIGMA-ALDRICH JAPAN K.K.
Tokyo Tel: 03 5796 7300
Tokyo Fax: 03 5796 7315
Australia
SIGMA-ALDRICH PTY., LIMITED
Free Tel: 1800 800 097
Free Fax: 1800 800 096
Tel: 612 9841 0555
Fax: 612 9841 0500
Germany
SIGMA-ALDRICH CHEMIE GmbH
Free Tel: 0800-51 55 000
Free Fax: 0800-649 00 00
Korea
SIGMA-ALDRICH KOREA
Tel: 031-329-9000
Fax: 031-329-9090
Austria
SIGMA-ALDRICH HANDELS GmbH
Tel: 43 1 605 81 10
Fax: 43 1 605 81 20
Belgium
SIGMA-ALDRICH NV/SA.
Free Tel: 0800-14747
Free Fax: 0800-14745
Tel: 03 899 13 01
Fax: 03 899 13 11
Brazil
SIGMA-ALDRICH BRASIL LTDA.
Tel: 55 11 3732-3100
Fax: 55 11 3733-5151
Canada
SIGMA-ALDRICH CANADA LTD.
Free Tel: 800-565-1400
Free Fax: 800-265-3858
Tel: 905-829-9500
Fax: 905-829-9292
China
SIGMA-ALDRICH CHINA INC.
Tel: 86-21-6386 2766
Fax: 86-21-6386 3966
Czech Republic
SIGMA-ALDRICH s.r.o.
Tel: 246 003 200
Fax: 246 003 291
Denmark
SIGMA-ALDRICH DENMARK A/S
Tel: 43 56 59 10
Fax: 43 56 59 05
Finland
SIGMA-ALDRICH FINLAND
Tel: 358-9-350-92 50
Fax: 358-9-350-92 555
Greece
SIGMA-ALDRICH (O.M.) LTD
Tel: 30 210 9948010
Fax: 30 210 9943831
Hungary
SIGMA-ALDRICH Kft
Tel: 06-1-235-9054
Fax: 06-1-269-6470
Ingyenes zld telefon: 06-80-355-355
Ingyenes zld fax: 06-80-344-344
India
SIGMA-ALDRICH CHEMICALS
PRIVATE LIMITED
Telephone
Bangalore: 91-80-5112-7272
Hyderabad:
91-40-5531 5548 / 2784 2378
Mumbai: 91-22-2579 7588 / 2570 2364
New Delhi:
91-11-2616 5477 / 2619 5360
Fax
Bangalore: 91-80-5112-7473
Hyderabad: 91-40-5531 5466
Mumbai: 91-22-2579 7589
New Delhi: 91-11-2616 5611
Malaysia
SIGMA-ALDRICH (M) SDN. BHD
Tel: 603-56353321
Fax: 603-56354116
Mexico
SIGMA-ALDRICH QUMICA, S.A. de C.V.
Free Tel: 01-800-007-5300
Free Fax: 01-800-712-9920
The Netherlands
SIGMA-ALDRICH CHEMIE BV
Tel Gratis: 0800-0229088
Fax Gratis: 0800-0229089
Tel: 078-6205411
Fax: 078-6205421
New Zealand
SIGMA-ALDRICH PTY., LIMITED
Free Tel: 0800 936 666
Free Fax: 0800 937 777
Norway
SIGMA-ALDRICH NORWAY AS
Tel: 23 17 60 60
Fax: 23 17 60 50
Ireland
SIGMA-ALDRICH IRELAND LTD.
Free Tel: 1800 200 888
Free Fax: 1800 600 222
Poland
SIGMA-ALDRICH Sp. z o.o.
Tel: (+61) 829 01 00
Fax: (+61) 829 01 20
Israel
SIGMA-ALDRICH ISRAEL LTD.
Tel: 08-948-4100
Fax: 08-948-4200
Portugal
SIGMA-ALDRICH QUMICA, S.A.
Free Tel: 800 20 21 80
Free Fax: 800 20 21 78
Italy
SIGMA-ALDRICH S.r.l.
Telefono: 02 33417310
Fax: 02 38010737
Numero Verde: 800-827018
Russia
SIGMA-ALDRICH RUSSIA
OOO SAF-LAB
Tel: 095 975-1917/3321
Fax: 095 975-4792
Singapore
SIGMA-ALDRICH PTE. LTD.
Tel: 65-6271 1089
Fax: 65-6271 1571
South Africa
SIGMA-ALDRICH
SOUTH AFRICA (PTY) LTD.
Tel: 27 11 979 1188
Fax: 27 11 979 1119
Spain
SIGMA-ALDRICH QUMICA S.A.
Free Tel: 900101376
Free Fax: 900102028
Sweden
SIGMA-ALDRICH SWEDEN AB
Tel: 020-350510
Fax: 020-352522
Outside Sweden Tel: +46 8 7424200
Outside Sweden Fax: +46 8 7424243
Switzerland
FLUKA CHEMIE GmbH
Swiss Free Call: 0800 80 00 80
Tel: +41 81 755 2828
Fax: +41 81 755 2815
United Kingdom
SIGMA-ALDRICH COMPANY LTD.
Free Tel: 0800 717181
Free Fax: 0800 378785
Tel: 01747 833000
Fax: 01747 833313
United States
SIGMA-ALDRICH
P.O. Box 14508
St. Louis, Missouri 63178
Toll-free: 800-325-3010
Call Collect: 314-771-5750
Toll-Free Fax: 800-325-5052
Tel: 314-771-5765
Fax: 314-771-5757
Internet:
sigma-aldrich.com
sigma-aldrich.com
Order/Customer Service 1-800-325-3010 Fax 1-800-325-5052
Technical Service 1-800-325-5832 sigma-aldrich.com/techservice
Development/Bulk Manufacturing Inquiries Sigma-Aldrich Fine Chemicals 1-800-336-9719
World Headquarters 3050 Spruce St., St. Louis, MO 63103 (314) 771-5765
We are committed to the success of our Customers, Employees and Shareholders through leadership in Life Science, High Technology and Service.
GDR
00573-501980
0024
2004 Sigma-Aldrich Co. Printed in USA. Sigma brand products are sold through Sigma-Aldrich, Inc.
Sigma-Aldrich, Inc. warrants that its products conform to the information contained in this and other Sigma-Aldrich publications. Purchaser must determine the suitability of the product(s) for their
particular use. Additional terms and conditions may apply. Please see reverse side of the invoice or packing slip. SIGMA and - are registered trademarks of Sigma-Aldrich Co. and its division
Sigma-Aldrich Biotechnology LP. Riedel-de Han: trademark under license from Riedel-de Han GmbH.