WB and tissue samples - (Sep/29/2006 )
...I have a question....I extracted protein from some mouse organs but since I just have an monoclonal antibody to detect my protein I always have really high background (I tried to block with all kind of agent, wash with different solution)....and I was thinking, that might be due to a cross reaction between IgG in the tissue and IgG from the antibody (infact I have the highest background in tissue like spleen and liver)...if this is true could help incubate my lisate with protein G beads (that bind IgG)?...anyone have a suggestion to give to me?
clearing for unspecific IgG binding with protein G or original secondary antibody + protein A/G or conjugated secondary Ab may improve your signal/noise ratio (by lowering the noise); you may also increase your signal by specific immunoprecipitation of your antigen out of the lysat; however, for probing on Wb, you need another Ab than for immunoprecipitation even if your Ab for IB is suitable for Wb
Thank you for the suggestion... I will try:ph34r: