Hello everyone,

I am struggling to find HDM-specific IgE in the sera of my allergy model. I tried several protocols for ELISA with changing the serum dilution (from 100x to 2x; also pre-clearing the sera with protein G), the incubation time of the sera (from 1-2h RT/37oC to ON at 4oC), and the enzyme reaction (AP, HRP, Fluorescent ELISA with HRP). I also changed the allergic model several times to find the best and I found HDM-specific IgG1 but not an IgE (Chronic model with only intranasal challenges and model with intraperitoneal sensitization with HDM + ALUM).

The HDM extract and Der p1-biotin that I use are from Citeq Biologics and the anti-IgE with or without biotin is a clone 23G3.

Please share your best tips for HDM-specific IgE ELISA, and suggest where I can be wrong with my protocol.

Thanks in advance!