Detection of Mouse Anti-OVA Specific IgG1 Serum Antibody

Microbiology and Immunology
Loyola University Chicago
Research Associate

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Company:

BioLegend

Product Name:

Anti-mouse IgG1 antibody

Catalog Number:

406604

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.

Experimental Design and Results Summary

Applications

ELISA

Sample

Serially diluted mouse sera was added in microplate precoated with 5ug/ml OVA and incubated for 1 hr at 37 degrees

Primary Incubation

Biotin labeled anti-mouse IgG1 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 was added (1:10,000) and incubated for 1 hr at 37 degrees

Tertiary Incubation

NA

Detection

TMB was used for signal development

Results Summary

We found that the level of ovalbumin (OVA)-specific IgG1 was significantly reduced in Trpm2−/− mice after immunization with OVA-encapsulated DOTAP liposomes. The antibody is sensitive and specific for ELISA.

Additional Notes

None

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Summary

The Good

This antibody is sensitive and specific for detecting antigen specific IgG1 antibody isotype.

The Bad

None

The Bottom Line

The antibody is able to specifically detect mouse IgG1 isotype antibody by ELISA. We do not see much non-specific reactions.

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