Order ISO-M8a: Each kit contains two cassettes, one for IgG1, IgG2a, IgG2b and IgG3, the other κ, λ, IgA, and IgM.
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Manual >>Rapid Mouse Immunoglobulin Isotyping Kit – ISO-M8a
Benefit:
The Rapid Mouse Immunoglobulin Isotyping Kit is a five-minute lateral flow assay with ELISA sensitivity for monoclonal antibody class and subclass determination. It can determine IgG1, IgG2a, IgG2b, IgG3, IgA, and IgM, as well as κ, λ in one quick test.
Acceptable Sample Types:
include hybridoma culture supernatant and purified monoclonal antibody. Ascites fluid is not recommended since it usually contains multiple immunoglobulins. However, this kit can still be used to determine the relative distribution of the main isotypes in a sample of ascites fluid.
Kit Components:
The isotyping kit has three different sizes with 5, 10 or 20 pouches containing two cassettes per pouch: one cassette is for determining IgG1, IgG2a, IgG2b and IgG3, and the other cassette is for determining κ, λ, IgA and IgM. Both strips contain a control line.
Note:
BALB/c and DBA mice produce IgG2a, not IgG2c. But C57BL/6, C57BL/10, SJL and NOD produce IgG2c, not 2a*.
Related Products:
Mouse Isotyping Kit M8c- Rapid Mouse Antibody Isotyping XpressCard (ISO-M8c): detect IgG2c in addition to IgG1, IgG2b, IgG3, IgA, and IgM, as well as κ, λ.
Mouse Isotyping Kit M8ac- Rapid Mouse Antibody Isotyping XpressCard (ISO-M8ac): IgG1, IgG2a, IgG2b, IgG2c, IgG3, IgM, κ and λ.
Mouse Isotyping Kit M6-Rapid Mouse Antibody Isotyping XpressCard (ISO-M6): determine IgG1, IgG2a, IgG2b, IgM, κ, and λ.
Protocol:
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.
This assay is faster and more convenient than the traditional ELISA method.
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: Besides the positive control line, more than one heavy chain and/or light chain will appear. The line intensity can be used to judge the dominant isotypes.
Performance :
Minimal detection limit is 100 ng/m. There is no cross-reactivity among each isotype.
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.
A Video Demonstration:
*1) The need for IgG2c specific antiserum when isotyping antibodies from C57BL/6 and NOD mice. Martin RM, et al (1998) J. Immunol Methods 212 (2): 187-192
2) Possible allelic structure of IgG2a and IgG2c in mice. Zhang Z et al (2012) Mol Immunol 50 (3): 169-171
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