RH FACTOR

1. The Rhesus factor, also known as the Rh factor, gets its name
from experiments conducted in 1937 by scientists Karl
Landsteiner and Alexander S. Weiner
2. The Rh factor is an antigen (a protein) that is found on the
surface of the red blood cell.
3. There are four general categories of blood: A, B, O, and AB. Each
blood type is further labeled as positive or negative, which is a
reference to the Rhesus factor of the blood
4. People with the Rhesus factor, that is, people with the antigen
present in their blood, are Rh-positive.
5. People without the Rhesus factor, that is, people that don't have
the antigen in their blood, are Rh-negative.
6. a Rh-positive child born to an Rh-negative woman runs the risk of
developing Rh disease
7. Only Rh-negative women risk having children with the Rhesus
factor disease; Rh-positive women do not.

HORSERADISH PEROXIDASE

1. The horse radish peroxidase enzyme (HRP) is used in the

DIAPOPS procedure to generate a color signal from a substrate
2. Superior to alkaline phosphatase and ß-galactosidase conjugates
due to the higher specific enzyme activity
3. Small molecular size (40,000 M.W.) allows excellent cellular
penetration
4. Variety of substrates available
5. Ideal in immunoblotting and immunocytochemistry applications
6. Used as the reporter enzyme for Super Signal Chemiluminescent
Substrates
7. Conjugates compatible with a number of substrates

AMYLASE

1. Amylase is an enzyme that helps digest carbohydrates

2. It is produced mainly in the pancreas and the glands that make
saliva
3. When the pancreas is diseased or inflamed, amylase releases into
the blood
4. The blood amylase test is ordered, often along with a lipase test,
to help diagnose and monitor acute or chronic pancreatitis and
other disorders that may involve the pancreas
 5. Amylase tests are sometimes used to monitor treatment of
cancers involving the pancreas and after the removal of
gallstones that have caused gallbladder attacks
6. Drug that can increase amylase measurements include:
Asparaginase, Aspirin, Birth control pills, etc.

SDS-PAGE

1. The purpose of this method is to separate proteins according to

their size, and no other physical feature
2. Proteins are denatured by SDS, the larger they are the more SDS
they bind and the more negatively charged they become
3. The electrophoresis of proteins through a polyacrylamide gel is
carried out in the presence of the detergent sodium dodecyl
sulfate (SDS).
4. SDS (sodium dodecyl sulfate) is a detergent (soap) that can
dissolve hydrophobic molecules but also has a negative charge
(sulfATE) attached to it
5. This method provides a relatively simple and highly effective
means of separating mixtures of proteins on the basis of size.
6. SDS is a negatively charged detergent that binds to protein in
amounts proportional to the length of the protein.
7. This binding destroys the characteristic tertiary and secondary
structure of the protein.
8. All proteins have a large negative charge which means they will
all migrate towards the positve pole when placed in an electric
field

IMMOBILIZED ENZYME

1. An immobilized enzyme is: an enzyme that is physically attached

to a solid support over which a substrate is passed and converted
to product.
2. This can provide increased resistance to changes in conditions
such as pH or temperature
3. An alternative to enzyme immobilization is whole cell
immobilization.
4. The surface on which the enzyme is immobilized is responsible
for retaining the structure in the enzyme through hydrogen
bonding or the formation of electron transition complexes
5. Method includes Carrier-Binding : the binding of enzymes to
water-insoluble carriers, Cross-Linking : intermolecular cross-
 linking of enzymes by bi-functional or multi-functional reagents,
Entrapping : incorporating enzymes into the lattices of a semi-
permeable gel or enclosing the enzymes in a semi-permeable
polymer membrane
6. Some applications are, Isomerization of glucose to fructose (High
fructose syrup), Lactose hydrolysis (dairy products converted to
avoid lactose intolerance), Penicillin acylase and Amino acid
synthesis

ROBERT KOCH

1. Robert Koch was born in 1843. Koch worked on anthrax and

tuberculosis (TB)
2. He was awarded the Nobel Prize in Physiology or Medicine for his
tuberculosis findings in 1905
3. He became famous for isolating Bacillus anthracis (1877), the
Tuberculosis bacillus (1882) and the Vibrio cholerae (1883) and
for his development of Koch's postulates
4. During this period Koch returned to his work on tuberculosis. He
sought to arrest the disease by means of a preparation, which he
called tuberculin, made from cultures of tubercle bacilli
5. Koch's pupils found the organisms responsible for diphtheria,
typhoid, pneumonia, gonorrhoea, cerebrospinal meningitis,
leprosy, bubonic plague, tetanus, and syphilis, among others, by
using his methods.
6. Robert Koch died on 27 May 1910 from a heart-attack in Baden-
Baden, aged 66.

WRIGHT'S STAIN

Wright's stain is a histologic stain that facilitates the differentiation of

blood cell types
It is named for James Homer Wright, who devised the stain, a
modification of the Romanowsky stain, in 1902
Wright's stain is the combination of acid dye (red 'eosin') and a basic
dye (blue 'methylene blue') for use of staining the blood smear
It highlights the differences among the different types of leukocytes
(immune cells) for easier recognition of eosinophils or basophils during
the counting process
There are various related staining products of Wright's stain which are
combinations of eosin, azure, and methylene blue in pH adjusted
solution
In cytogenetics it is used to stain chromosomes to facilitate diagnosis
of syndromes and diseases

LEISHMAN'S STAIN

1. A neutral stain for blood smears devised by the British surgeon

W. B. Leishman (1865–1926).
2. Leishman's stain, also Leishman stain, is used in microscopy
for staining blood smears
3. It consists of a mixture of eosin (an acidic stain), and methylene
blue (a basic stain) in alcohol and is usually diluted and buffered
before use
4. It stains the different components of blood in a range of shades
between red and blue.
5. It is generally used to differentiate and identify leucocytes,
malaria parasites, and trypanosomas
6. It is similar to and partially replaceable with Giemsa stain,
Jenner's stain, and identical to Wright's stain.

PRECIPITIN BAND

1. Precipitin band can be detected when using very high

concentration of protein (100 µ g/ml)
2. Precipitin band get disrupt when more antigen diffuses.
3. The original precipitin reaction was carried out by layering a
solution of antibody on top of a solution of stain extract in a tube,
and left for a period of time to allow the development of a
precipitin band at the interface
4. A precipitin band is formed when the diffusing stain contains
proteins that are recognized by IgG molecules in the diffusing
antiserum.
5. The precipitin band is sometimes clearly visible to the naked eye.
6. But it is normal to stain the plates with amino black or other
general protein stain, to enhance sensitivity and clarity.

SKIM MILK AGAR

1. Skim Milk is used for preparing microbiological culture media.

2. Skim Milk Medium may be used for the cultivation and
differentiation of microorganisms based on the coagulation and
proteolysis of casein.
3. Skim Milk can be used to prepare Skim Milk Agar for detecting
proteolytic microorganisms in foods, including dairy products
 4. Skim Milk is a source of lactose and casein and other nutrients
required for the growth of lactobacilli.
5. Clostridial species can be differentiated based on their ability to
enzymatically degrade proteins to peptones (peptonization) or
coagulate milk.
6. It may be used to detect the stormy fermentation produced by
Clostridium perfringens.
7. Addition of skim milk to a culture medium with a superior nutrient
base optimally adapts it to the neutral conditions experienced by
microorganisms which grow in milk.

LOUIS PASTEUR

1. Louis Pasteur was a world renowned French chemist and biologist

born on December 27 1822
2. Pasteur founded the science of microbiology and proved that
most infectious diseases are caused by micro-organisms. This
became known as the "germ theory" of disease.
3. He was the inventor of the process of pasteurisation and also
developed vaccines for several diseases including rabies.
4. He solved the mysteries of rabies, anthrax, chicken cholera, and
silkworm diseases, and contributed to the development of the
first vaccines. He debunked the widely accepted myth of
spontaneous generation
5. The discovery of the vaccine for rabies led to the founding of the
Pasteur Institute in Paris in 1888.
6. On July 6 1885, Pasteur tested his pioneering rabies vaccine on
man for the first time
7. He died at Saint-Cloud on 28 September 1895.

THYMUS

1. The name thymus comes from a supposed resemblance to the

bud of the herb thyme (in Latin, thymus).
2. A small glandular organ that is situated behind the top of the
breastbone
3. It consisting mainly of lymphatic tissue and serving as the site of
T cell differentiation
4. The thymus increases gradually in size and activity until puberty,
becoming vestigial thereafter.
 5. The generation of T-cells expressing distinct T-cell receptors
occurs within the thymus
6. As the thymus is the organ of T-cell development, any congenital
defect in thymic genesis or a defect in thymocyte development
can lead to a profound T cell primary immunodeficiency.

IMMUNOGLOBULIN A

1. Immunoglobulin A (IgA) is an antibody that plays a critical role in

mucosal immunity
2. IgA has two subclasses (IgA1 and IgA2) and can exist in a dimeric
form called secretory IgA (sIgA)
3. IgA is a poor activator of the complement system, and opsonises
only weakly. Its heavy chains are of the type α.
4. In the blood, IgA interacts with an Fc receptor called FcαRI (or
CD89), which is expressed on immune effector cells, to initiate
inflammatory reactions
5. Decreased or absent IgA, termed selective IgA deficiency, can be
a clinically significant immunodeficiency.
6. IgA nephropathy is caused by IgA deposits in the kidneys.

PCR

1. Developed in 1983 by Kary Mullis

2. The polymerase chain reaction (PCR) is a scientific technique
in molecular biology to amplify a single or a few copies of a piece
of DNA and is used to amplify a specific region of a DNA strand
(the DNA target)
3. The PCR process can be done by various steps: Initialization step,
Denaturation step, Annealing step, Extension/elongation step,
Final elongation and Final hold
4. PCR allows isolation of DNA fragments from genomic DNA by
selective amplification of a specific region of DNA
5. PCR permits early diagnosis of malignant diseases such as
leukemia and lymphomas, which is currently the highest-
developed in cancer research and is already being used routinely
6. PCR also permits identification of non-cultivatable or slow-
growing microorganisms such as mycobacteria, anaerobic
bacteria, or viruses from tissue culture assays and animal
models.
AGAROSE

1. Agarose is a polysaccharide obtained from agar that is used for a

variety of life science applications especially in gel
electrophoresis
2. It accumulates in the cell walls of agarophyte red algae.
3. Agarose is a linear polymer, made up of the repeating monomeric
unit of agarobiose
4. Agarose forms an inert matrix utilized in separation techniques
5. Many structures easily affix to agarose including various types of
proteins
6. Agar (agar-agar) can be used as a laxative, a vegetarian gelatin
substitute, a thickener for soups, in jellies, ice cream and other
desserts, as a clarifying agent in brewing, and for sizing paper
and fabrics.

IMMUNODIFFUSION

1. Ouchterlony double diffusion (ODD) or immunodiffusion

technique is one of the simplest techniques extensively used to
check antisera for the presence of antibodies for a particular Ag
and to determine its titer.
2. A gel plate is cut to form a series of holes (wells) in the gel. A
sample extract of interest is placed in one well
3. Precipitation occurs with most antigens because the antigen is
multivalent (i.e. has several antigenic determinants per molecule
to which antibodies can bind)
4. Antibodies have at least two antigen binding sites
5. Amount protein precipitated increases until the antigen/antibody
molecules are at an optimal ratio. This is known as the zone of
equivalence or equivalence point
6. When the amount of antigen in solution exceeds the amount of
antibody, the amount of precipitation will decrease. This is known
as the antigen excess zone.

ELISA READER

1. Elisa Readers are laboratory instruments designed to detect

biological, chemical or physical events of samples in microtiter
plates
2. They are widely used in research, drug discovery, bioassay
validation, quality control and manufacturing processes in the
 pharmaceutical and biotechnological industry and academic
organizations.
3. Common detection modes for microplate assays are absorbance,
fluorescence intensity, luminescence, time-resolved fluorescence,
and fluorescence polarization.
4. It is used for assays such as ELISA assays, protein and nucleic
acid quantification or enzyme activity assays
5. The range of applications for multi-mode plate readers is
extremely large: Protein and cell growth assays, Enzyme activity,
Molecular interactions, Nucleic acid quantitation, Cell toxicity,
proliferation, and viability, etc.,

SERUM

1. Blood serum, a component of blood which is collected after

coagulation
2. The clear liquid that can be separated from clotted blood
3. Serum differs from plasma, the liquid portion of normal unclotted
blood containing the red and white cells and platelets
4. It is the clot that makes the difference between serum and
plasma.
5. Blood serum from the tissues of immunized animals, containing
antibodies and used to transfer immunity to another individual.
6. Blood serum contains 6–8% solids, including macromolecules
such as albumin, antibodies and other globulins, and enzymes;
peptide and lipid-based hormones; and cytokines; as well as
certain nutritive organic materials in small amounts, such as
amino acids, glucose, and fats.

BLOOD CELLS

1. A blood cell, also called a hematocyte, is a cell of any type

normally found in blood.
2. In mammals, these fall into three general categories: red blood
cells — Erythrocytes, white blood cells — Leukocytes and
platelets — Thrombocytes
3. Three kinds of granulocytes: neutrophils, eosinophils and
basophils
4. Two kinds of leukocytes without granules in their cytoplasm
lymphocytes , monocytes
 5. All the various types of blood cells are produced in the bone
marrow
6. All arise from a single type of cell called a hematopoietic stem
cell
7. Red blood cells have surface antigens that differ between people
and that create the so-called blood groups such as the ABO
system and the Rh system

ROCKET IMMUNOELECTROPHORESIS

1. Rocket electrophoresis (also referred to as electroimmunoassay

or electroimmunodiffusion) is a simple, quick, and reproducible
method for determining the concentration of a specific protein in
a protein mixture.
2. The method, originally introduced by Laurell
3. It is one-dimensional quantitative immunoelectrophoresis
4. It is used to determination of various concentration of antigen in
given unknown sample.
5. Various concentrations of antigen are loaded side by side in small
circular wells along the edge of an agarose gel that contains the
specific antibody (Ab)
6. On electrophoresis, the antigen begins to migrate towards the
anode and interacts with antibody molecules to form a soluble
antigen-antibody complex
7. The precipitin line is seen in the form of a rocket.

IgM

1. IgM antibodies are the largest antibody

2. They are found in blood and lymph fluid and are the first type of
antibody made in response to an infection
3. They also cause other immune system cells to destroy foreign
substances
4. IgM antibodies are about 5% to 10% of all the antibodies in the
body.
5. IgM for short, is a basic antibody that is produced by B cells
6. IgM in normal serum is often found to bind to specific antigens,
even in the absence of prior immunization
 7. IgM forms polymers where multiple immunoglobulins are
covalently linked together with disulfide bonds, mostly as a
pentamer but also as a hexamer.

RADIAL IMMUNODIFFUSION

1. Radial immunodiffusion (or Mancini method, Mancini

immunodiffusion, single radial immunodiffusion assay) is an
immunodiffusion technique used in immunology to determine the
quantity of an antigen by measuring the diameters of circles of
precipitin complexes
2. A quantitative immunodiffusion technique used to detect the
level of protein (antigen) in a sample by measuring the diameter
of the ring of precipitin formed by the complex of the protein
(antigen) and the antiserum (antibody).
3. The diameters of the circles increase with time as the antigen
diffuses into the medium, reacts with the antibody, and forms
insoluble precipitin complexes.
4. Compared with microbial diffusion assays, this technique is less
precise as it gives a rather relatively insensitive estimation of the
concentration of antigen
5. It can be applied when sensitivity is not required but specificity is,
and antibody is plentiful.
6. It is commonly used in clinically detecting levels of blood proteins
in a patient

LYMPHOCYTES

1. A small white blood cell (leukocyte) that plays a large role in

defending the body against disease.
2. Lymphocytes are responsible for immune responses
3. There are two main types of lymphocytes: B cells and T cells.
4. The B cells make antibodies that attack bacteria and toxins
5. T cells attack body cells themselves when they have been taken
over by viruses or have become cancerous.
6. Lymphocytes secrete products (lymphokines) that modulate the
functional activities of many other types of cells and are often
present at sites of chronic inflammation.
RHEUMATOID FACTOR (RF)
1. A rheumatoid factor (RF) blood test measures the amount of the
RF antibody present in the blood
2. Normally, antibodies are produced by the immune system to help
destroy and eliminate invading bacteria and viruses that can
cause disease
3. But the RF antibody can attach to normal body tissue, resulting in
damage.
4. A high level of rheumatoid factor can be caused by several
autoimmune diseases (including rheumatoid arthritis) and some
infections.
5. Occasionally an elevated level of RF is present in healthy people.
6. A test for rheumatoid factor is done to help support a diagnosis of
rheumatoid arthritis.

HUMAN CHORIONIC GONADOTROPIN

1. A human hormone made by chorionic cells in the fetal part of the

placenta.
2. Human chorionic gonadotropin (hCG) is directed at the gonads
and stimulates them. Hence, the name "gonadotropin."
3. The presence of hCG is detectable by immunologic means within
days of fertilization and forms the foundation of the common
pregnancy tests.
4. The level of hCG tends to be higher with a female fetus soon after
conception.
5. The level of hCG in maternal serum also enters as one
component in the "double" and the "triple" screening tests used
during pregnancy to assign risks of Down syndrome and other
fetal disorders.
6. Human chorionic gonadotropin interacts with the LHCG receptor
and promotes the maintenance of the corpus luteum during the
beginning of pregnancy, causing it to secrete the hormone
progesterone
7. Levels of hCG may be measured in the blood or urine

IMMUNOELECTROPHORESIS

1. Immunoelectrophoresis, also called gamma globulin

electrophoresis, or immunoglobulin electrophoresis.
 2. It is a method of determining the blood levels of three major
immunoglobulins: immunoglobulin M (IgM), immunoglobulin G
(IgG), and immunoglobulin A (IgA).
3. Immunoelectrophoresis is a powerful analytical technique with
high resolving power as it combines separation of antigens by
electrophoresis with immunodiffusion against an antiserum.
4. It is usually requested when a different type of electrophoresis,
called a serum protein electrophoresis, has indicated a rise at the
immunoglobulin level.
5. Immunoelectrophoresis is also used frequently to diagnose
multiple myeloma, a disease affecting the bone marrow.
6. Drugs that may cause increased immunoglobulin levels include
therapeutic gamma globulin, hydralazine, isoniazid, phenytoin
(Dilantin), procainamide, oral contraceptives, methadone,
steroids, and tetanus toxoid and antitoxin.

WESTERN BLOT

1. The Western blot (alternatively, protein immunoblot) is a widely

used analytical technique used to detect specific proteins in the
given sample of tissue homogenate or extract.
2. A technique developed in 1979 that is used to confirm ELISA
results.
3. It uses gel electrophoresis to separate native or denatured
proteins by the length of the polypeptide (denaturing conditions)
or by the 3-D structure of the protein (native/ non-denaturing
conditions).
4. The proteins are then transferred to a membrane (typically
nitrocellulose or PVDF), where they are probed (detected) using
antibodies specific to the target protein.
5. bacteria, virus or environmental samples can be the source of
protein and thus Western blotting is not restricted to cellular
studies only
6. The confirmatory HIV test employs a Western blot to detect anti-
HIV antibody in a human serum sample

EDTA

1. The compound was first described in 1935 by Ferdinand Munz,

who prepared the compound from ethylenediamine and
chloroacetic acid
2. Ethylenediaminetetraacetic acid, widely abbreviated as EDTA
(for other names, see Table) is a polyamino carboxylic acid and a
colourless, water-soluble solid
3. Its conjugate base is named ethylenediaminetetraacetate
4. It is widely used to dissolve limescale
5. Its usefulness arises because of its role as a hexadentate ("six-
toothed") ligand and chelating agent
6. After being bound by EDTA, metal ions remain in solution but
exhibit diminished reactivity
7. EDTA is produced as several salts, notably disodium EDTA and
calcium disodium EDTA.