I am looking to measure the concentration of antigen-specific IgE antibodies in B6 mice (immunized with an antigen) using an ELISA, but I don't know if that will be possible. I know that the majority of IgE antibodies are bound to mast cells, so I'm not sure if I'll be able to detect any. Does anyone know of a method I could use to measure antigen-specific IgE concentrations?
I do ELISA, regularly to check the Ag specific IgG, but I donot whether same protocol (coating the Ag to the wells) will work for you or not. I found these two papers for you, these may be useful.
http://www.sciencedirect.com/science/article/pii/0022175989902020
http://www.ncbi.nlm.nih.gov/pubmed/20951440
You can also find ready to use (or precoated) ELISA kits.
Hi Ashish, there should be plenty of IgE present in the serum for you to measure, assuming your mice have made a sufficient IgE-mediated response to your antigen. We routinely measure peanut-specific IgE in mice that have been sensitized to peanut and get good levels of response by ELISA. We use a similar technique to Manoj - ie we coat ELISA plates first with peanut, then incubate with our sera and follow with an anti-IgE antibody.
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