Can I IP from media? - to use for SDS-PAGE? (Jun/09/2010 )
I know that my protein of interest is expressed on the cell membrane, and it is predicted to have a cleavage site in the extracellular domain.
I want to see if my protein is being cleaved and floating around in the media.
I have an antibody that will bind to the extracellular section.
Can I use this antibody to IP the protein from the media like I would from cell lysates or will that not work due to the large volume? Or do I need to do a TCA precipitation or something similar to concentrate the media first?
yes, u can, but it is best to concentrate the media using concentrator before IP, the medium may be too diluted.