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Anti-mouse IgG2b antibody
406704
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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.
ELISA
Serially diluted mouse serum was added on a microplate precoated with 5ug/ml OVA and incubated for 1 hr at 37 degrees.
Biotin labeled anti-mouse IgG2b antibody (0.5 ug/ml) was added and incubated for 1 hr at 37 degrees.
5% nonfat milk in PBST was used for plate blocking after OVA protein coating.
HPR-streptavidin were added (1:10,000) and incubated for 1 hr at 37 degrees
TMB was used for signal development
Plate was analyzed by ELISA plate reader for absorption at 450 nm
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.
None
Antibody is sensitive and has no cross-reactions
The antibody is able to specifically detect mouse IgG2b isotype antibody when used in ELISA. It has minimal non-specific reactions.