Protocol Online logo
Top : Forum Archives: : Immunology and Histology

URGENT:  IgG subclass - (Oct/14/2000 )

Can you please tell me an experimental design that would determine the subclass of a monoclonal Ab (IgG) given the following info: goat-anti-IgG3, goat-anti-IgG2a, goat-anti-IgG2b, goat-anti-IgG1...all @ 1:100. What if I use mAb being tested @ 1ug/ml? And also use goatXmouse IgG HRP conjugate @ 1:10,000?

Thank you. Your help would truly be appreciated.


Hi!At fisrt you need to coat the ELISA plate with the Antigent against which u have raised MAB.After blocking the unbound sites incubate with the MAB whose isotype you need to find.AFter that incubate with antimouse rabbit isotype detecting antibodies followed by incubation with enzyme conjugated anti rabbit goat antibody. Proceed for color detection as usual.

Good luckMK adde