a. Store the reagent at 2–8°C. Do not freeze.
b. The shelf-life of the reagent is as per the expiry date
mentioned on the reagent vial label.
Human red blood cells possessing A and/or B antigen will
agglutinate in the presence of antibody directed towards
the antigen. Agglutination of red blood cells with anti-A,
anti-B and anti-AB reagent is a positive test result and
indicates the presence of the corresponding antigen.
Absence of agglutination of red blood cells with anti-A,
anti-B and anti-AB reagent is a negative test result and
indicates the absence of corresponding antigen.
1. In vitro diagnostic reagent for laboratory and professional use only. Not for medicinal use.
2. Erybank reagents are from human source and the
source material used in its manufacture is tested by
approved techniques and found negative for HBsAg
3. The reagent contains sodium azide 0.1% as preservative.
Avoid contact with skin and mucosa. On disposal, flush
with large quantities of water.
4. Extreme turbidity may indicate microbial contamination
or denaturation of protein due to thermal damage.
Such reagent should be discarded.
Sample Collection and Preparation
No special preparation of the patient is required prior
to sample collection by approved techniques. Samples
should be stored at 2-8°C if not tested immediately. Do
not use hemolyzed samples. Anticoagulated blood using
various anticoagulants should be tested within the below
Sodium citrate or sodium oxalate : 14 days
Clotted whole blood should be tested within 14 days.
Additional Material Required for Slide and Tube
Glass slide (50 × 75 mm), test tubes (10 × 75 mm), Pasteur
pipettes, isotonic saline, centrifuge, timer, and mixing
Bring reagent and samples to room temperature before
1. Place one drop of Erybank reagent anti-A or anti-B or
anti-AB on a clean glass slide.
2. To each reagent drop, add one small drop of whole
3. Mix well with a mixing stick uniformly over an area of
4. Rock the slide gently back and forth.
5. Observe for agglutination macroscopically at 2
1. Prepare a 2–3% suspension of the red cells to be tested
2. Place one drop of Erybank anti-A, anti-B, anti-AB into
correspondingly labeled test tubes.
3. Pipette into each of the test tubes, one drop of the test
red cell suspension and mix well.
4. Centrifuge for 1 minute at 1000 rpm (125 g) or 20 seconds
at 3400 rpm (1000 g) or incubate at room temperature
Blood Banking (Immunohematology) 321
5. Gently resuspend the cell button, observing for agglutination macroscopically.
Agglutination is a positive test result and indicates the
presence of A and/or B antigen.
Do not interpret peripheral drying or fibrin strands as
No agglutination is a negative test result and indicates
the absence of A and/or B antigen.
1. In the tube test procedure, it is recommended that
tubes with negative reactions should be recentrifuged
and results read again after 5 minutes so that weak
2. Anti-A, lectin can be used to differentiate between A,
3. A3 cells demonstrate a mixed field agglutination.
4. Certain weaker subgroups of A and B may produce
weak reactions with anti-A, B.
5. As undercentrifugation or overcentrifugation could
lead to erroneous results, it is recommended that each
laboratory calibrate its own equipment and determine
the time required for achieving the desired results.
Monoclonal Blood Grouping Antibodies for
(Courtesy: Tulip’s Eryclone Range)
Monoclonal antibodies are derived from hybridoma cell
lines, created by fusing mouse antibody producing B
lymphocytes with mouse myeloma cells. Each hybridoma
cell line produces homogeneous antibodies of only
one immunoglobulin class, which are identical in their
chemical structure and immunological activity. Human
red blood cell antigens can be divided into four groups
A, B, AB and O depending on the presence or absence
of the corresponding antigens on the red blood cells.
Approximately, 41% of the Caucasian population have the
A antigen, 9% have the B antigen, 4% have both A and B
antigens, while the remaining have neither A nor B antigen.
Eryclone® anti-A, anti-B and anti-AB are ready-to-use
reagent prepared from supernatants of mouse hybridoma
cell cultures. These antibodies of immunoglobulin class
IgM are a mixture of several monoclonal antibodies of the
same specificity but having the capability of recognizing
different epitopes of the human red blood cell antigens
A and B. Each batch of reagent undergoes quality control
at various stages of manufacture for its specificity, avidity
1. Store the reagent at 2–8°C. Do not freeze.
2. The shelf life of reagent is as per the expiry date mentioned on the reagent vial label.
Human red blood cells possessing A and/or B antigen will
agglutinate in the presence of antibody directed towards
the antigen. Agglutination of red blood cells with anti-A,
anti-B, anti-AB reagent is a positive test result and indicates
the presence of the corresponding antigen. Absence of
agglutination of red blood cells with anti-A, anti-B, anti-AB
reagent is a negative test result and indicates the absence
1. In vitro diagnostic reagent for laboratory and professional use only. Not for medicinal use.
2. The reagent contains sodium azide 0.1% as preservative. Avoid contact with skin and mucosa. On
disposal, flush with large quantities of water.
3. Extreme turbidity may indicate microbial contamination or denaturation of protein due to thermal
damage. Such reagent should be discarded.
4. Eryclone reagent are not from human sources, hence,
contamination due to HBsAg and HIV is practically
Sample Collection and Preparation
No special preparation of the patient is required prior
to sample collection by approved techniques. Samples
should be stored at 2–8°C if not tested immediately. Do
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