i have a question for IP and antibody heavy and light chain detection.
I performed an Immunoprecipitation experiment but when i ran the SDS-PAGE gel, i loaded the rabbit antibody used for ip in a control lane however
doing the western blot with Goat anti-rabbit secondary antibody only detected a very strong 50kDa band in the lane!
Did it mean that the antibody is not completely denatured?
Should i expect another 20-30 kDa band on the gel corresponding to the antibody light chain?
I also boiled the antibody in loading buffer for 5 min so shouldnt this be ok?
Thanks again in advance