Detection of Mouse Anti OVA Specific IgG2b Antibody

Microbiology and Immunology
Loyola University Chicago
Research Associate

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Company:

BioLegend

Product Name:

Anti-mouse IgG2b antibody

Catalog Number:

406704

Info: View Product View Product Specs

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To test antibody responses of TRPM2 -/- mice, wild-type (Trpm2+/+) or Trpm2−/− mice were subcutaneously immunized on day 0 and day 14 with 40 μg/mouse ovalbumin (OVA) alone or the same amount of OVA encapsulated within DOTAP liposomes. On day 24, the mice were sacrificed, and sera were collected and analyzed for OVA-specific IgG1, IgG2b by ELISA. We found IgG1 level was reduced in OVA-DOTAP immunized mice, while IgG2b level did not change in either wild type of Trpm2-/- mice.

Experimental Design and Results Summary

Applications

ELISA

Sample

Serially diluted mouse serum was added on a microplate precoated with 5ug/ml OVA and incubated for 1 hr at 37 degrees.

Primary Incubation

Biotin labeled anti-mouse IgG2b antibody (0.5 ug/ml) was added and incubated for 1 hr at 37 degrees.

Blocking Agent

5% nonfat milk in PBST was used for plate blocking after OVA protein coating.

Secondary Incubation

HPR-streptavidin were added (1:10,000) and incubated for 1 hr at 37 degrees

Tertiary Incubation

TMB was used for signal development

Detection

Plate was analyzed by ELISA plate reader for absorption at 450 nm

Results Summary

We found that the level of ovalbumin (OVA)-specific IgG1 was reduced in Trpm2−/− mice after immunization with OVA-encapsulated DOTAP liposomes, but IgG2b level did not change in either wild type of Trpm2-/- mice.

Additional Notes

None

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Summary

The Good

Antibody is sensitive and has no cross-reactions

The Bad

None

The Bottom Line

The antibody is able to specifically detect mouse IgG2b isotype antibody when used in ELISA. It has minimal non-specific reactions.

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