Protein-A HRP to reduce background from HC/LC - (Oct/17/2007 )

Hi, I have been trying to optimize conditions to use Protein A-HRP conjugate to reduce background contamination from Heavy and light chains. Does anybody have a good protocol for using it. Also what is the typical secondary antibody concentration needed.

Any input is appreciated. Thanks.

you should run reduced Ig on a gel, transfer, cut the lanes and try several dilutions of protein A-HRP to find the one that do not detect the reduced Ig (HC and LC).
Depending on the batch and the provider, you can try from 5,000 to 200,000 I guess