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SDS­PAGE

Sodium Dodecyl Sulfate ­ 
Polyacrylamid Gel 
Electrophoresis
Sodium Dodecyl Sulfate
● SDS is a common ingredient in detergents
● Other names for SDS include laurel sulfate and
sodium laurel sulfate
● As a detergent SDS destroys protein secondary,
tertiary and quaternary structure
● This makes proteins rod shaped
● SDS also sticks to proteins in a ratio of
approximately 1.4 g of SDS for each gram of
protein
● Negative charge on the sulfate groups of SDS
mask any charge on the protein
SDS
Sodium Dodecyl Sulfate
C12H25NaO4S
H H H H H H H H H H H H O
H-C-C-C-C-C-C-C-C-C-C-C-C-O-S-O-Na+
H H H H H H H H H H H H O
Non-polar Polar
Hydrophobic tail Hydrophilic head

● Because it is amphipathic, SDS is a potent detergent


● SDS may also be called sodium lauryl sulfate and is a
common ingredient of shampoos laundry detergent and
other cleaning products
SDS and Proteins

SDS

Protein
SDS and Proteins
● SDS nonpolar chains arrange themselves on proteins and
destroy secondary tertiary and quarternary structrure
● Thus shape is no longer an issue as the protein SDS
complex becomes rod shaped

● In aqueous solutions, SDS polarizes releasing Na+ and


retaining a negative charge on the sulfate head
● So much SDS binds to proteins that the negative charge
on the SDS drowns out any net charge on protein side
chains
● In the presence of SDS all proteins have uniform shape
and charge per unit length
Polyacrylamide Gels
● Polyacrilamide is a polymer made of acrylamide
(C3H5NO) and bis-acrilamide (N,N’-methylene-
bis-acrylamide C7H10N2O2)

CH2 CH Acrylamide
O
O C NH2
C NH2
CH2
CH2 CH
O C NH2
Acrylamide
Acrylamide CH2 CH

bis-Acrylamide
Polyacrylamide Gels
● Acrylamide polymerizes in the presence of free
radicals typically supplied by ammonium persulfate

O O

C NH2 C NH2
CH2 CH CH2 CH

SO4-.
Polyacrylamide Gels
● Acrylamide polymerizes in the presence of free
radicals typically supplied by ammonium persulfate
● TMED (N,N,N’,N’-tetramethylethylenediamine)
serves as a catalyst in the reaction
O O O O

C NH2 C NH2 C NH2 C NH2


CH2 CH CH2 CH CH2 CH CH2 CH

SO4-.
Polyacrylamide Gels
● bis-Acrylamide polymerizes along with acrylamide
forming cross-links between acrylamide chains
O O O
C NH2 C NH2 C NH2
CH2 CH CH2 CH CH2 CH CH2 CH

O C NH2
CH2 O O O
O C NH2 C NH2 C NH2 C NH2
CH2 CH CH2 CH CH2 CH CH2 CH
bis-Acrylamide
Polyacrylamide Gels
● bis-Acrylamide polymerizes along with acrylamide
forming cross-links between acrylamide chains
Polyacrylamide Gels
● Pore size in gels can be varied by varying the ratio
of acrylamide to bis-acrylamide
● Protein separations typically use a 29:1 or 37.5:1
acrylamide to bis ratio

Little
Lots ofbis-acrylamide
bis-acrylamide
SDS­PAGE
1 2 3

3
Addition
2 of SDS
1 Protein becomes rod-
shaped with uniform
charge distribution