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The rapid mouse antibody isotyping test kit to determine the isotype of mouse IgG1, IgG2a, IgG2b, IgG3, IgA, IgM, kappa, and lambda in one test kit. Rapid, convenient, and accurate. Antibody isotyping test kits including mouse IgG2c, human, rat, and hamster are available.
Price/availability/specifications subject to change without notice. Unless otherwise indicated, our catalog and customized products are for research use only and not intended for human or animal diagnostic or therapeutic use.
Phone: 1-617-401-8149
Fax: 1-617-606-5019
Email: message@sydlabs.com
Or leave a message with a formal purchase
order (PO) Or credit card.
Background
A typical immunoglobulin (antibody) molecule is composed of four polypeptide chains, two identical heavy chains and two identical light chains. In mice, based on the heavy chain constant regions, antibodies are classified into IgA, IgD, IgE, IgG, and IgM, and the IgG isotype is further divided into IgG1, IgG2a (or IgG2c), IgG2b, and IgG3; the light chain constant regions are classified into kappa (κ) and lambda (λ). The IgG isotypes share more than 95% homology in the amino acid sequences of the Fc regions but show major differences in the amino acid composition and structure of the hinge region. Murine kappa/lambda ratio is 95/5, significantly different from the human kappa/lambda ratio of 60/40.
Naive B cells produce IgM and IgD; mature B cells produce one of IgG, IgA, and IgE through isotypic switching.
The isotypes of mouse IgG antibody reagents used in scientific research and clinical diagnostics are mainly mouse IgG1 kappa. Mouse IgG1 antibodies and engineered mutants are also used in therapeutic drug discovery. Mouse IgG1 antibodies are more like human IgG4 antibodies than human IgG1 antibodies, which behave more like mouse IgG2a or IgG2c antibodies. Many antibody drugs especially those targetting immune checkpoint proteins use the human IgG4 constant regions. The mouse IgG2c antibody is produced in certain inbred strains such as C57BL/6, C57BL/10, SJL, and NOD, which does not express the mouse IgG2a antibody, and vice versa.
The isotype and subclass for a mouse antibody can be determined by specific anti-heavy or light chain constant region antibodies, usually through lateral flow in-vitro immunoassay or ELISA, with appropriate isotype controls. We have developed rapid antibody isotyping test kits to determine the isotypes of antibody heavy and light chains as rapidly, conveniently, and accurately as many pregnancy test kits.
The rapid mouse antibody isotyping test kit A is a five-minute lateral flow assay with ELISA sensitivity for monoclonal antibody class and subclass determination. It can be used to determine the isotype of IgG1, IgG2a, IgG2b, IgG3, IgA, and IgM, as well as kappa and lambda in one test kit. There are two cassettes in the test kit, one for IgG1, IgG2a, IgG2b, and IgG3, and the other for kappa, Lamda, IgA, and IgM.
Sensitivity: Minimal detection limit is 100 ng/ml.
Specificity: There is no cross-reactivity among each isotype.
Sample requirements: hybridoma culture supernatant and purified monoclonal antibody. Although ascites fluid is not recommended since there is usually multiple immunoglobulins, the kit can be used to determine the relative distribution of the main isotypes in the ascites fluid.
Note: BALB/c and DBA mice produce IgG2a, not IgG2c. And C57BL/6, C57BL/10, SJL and NOD mice produce IgG2c, not IgG2a.
Sample preparation:
Cell culture supernatant: 1:10~1:100 diluted in PBS.
Purified Antibody: 0.5 – 2 µg/mL in PBS.
Ascites fluid: 1:10,000 diluted in PBS.
Protocol of the rapid mouse antibody isotyping test kit A:
1) Place the cassettes in horizontal plane.
2) Add 80-100 µL antibody solution directly to the sample well. Wait for 5 minutes.
3) A red test line at the location corresponding to antibody-specific isotype and a red control line at “C" position will appear within 1-5 minutes.
Result interpretation:
1. Invalid result: no red line at “C”.
2. Negative result: only one red line at “C”.
3. Positive results:
a). Single monoclonal antibody: In addition to the positive control “C” line, only one heavy chain and one light chain will appear.
b). Multiple monoclonal antibodies: In addition to the positive control “C” line, more than one heavy chain and/or light chain will appear. The line intensity can be used to determine the dominant isotypes.
Shipping: The rapid mouse antibody isotyping test kit A is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage: Store the strips in their original package at room temperature. This product is stable for up to 18 months from the certified date. Freezing the strips is not recommended.
Other rapid mouse antibody isotyping test kits:
Rapid Mouse Monoclonal Antibody Isotyping Test Kit C: IgG1, IgG2c, IgG2b, IgG3, IgA, IgM, kappa, and lambda.
Rapid Mouse Monoclonal Antibody Isotyping Test Kit A+C: IgG1, IgG2a, IgG2b, IgG2c, IgG3, IgA, IgM, kappa, and lambda.
Other rapid antibody isotyping test kits:
Rapid Human Monoclonal Antibody Isotyping Test Kit
Rapid Rat Monoclonal Antibody Isotyping Test Kit
Rapid Hamster Monoclonal Antibody Isotyping Test Kit
Recombinant mouse IgG isotype control antibodies:
In vivo Grade Recombinant Mouse IgG1 Isotype Control Antibody and Mutants
In vivo Grade Recombinant Mouse IgG2a Isotype Control Antibody and Mutants
In vivo Grade Recombinant Mouse IgG2b Isotype Control Antibody and Mutants
In vivo Grade Recombinant Mouse IgG2c Isotype Control Antibody and Mutants
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