aspecific band of BSA in western blot - western blotting (Oct/20/2003 )

I am working with bovine material, which has high BSA levels. Now I want to perform Western Blots for different matrixmetalloproteinases which have MW, such as 72, 68, and 52kDa. I always have strong aspecific bands for BSA with the blocking agents that I have used so far (5% dry milk, 1.5% gelatin, 3% goat serum, all in 0.1% tween TBS). Using lesser % of SDS gels (e.g. 10 or 8%) gives some more space between the bands, but I really would like to get rid of the BSA signal , because it may interfere with the bands I am interested in.

Does anyone now about this problem and have a solution?

Vidya

Hi there!

I've almost the same problem -high BSA levels in the medium I have to use- and want to analyse the sekretion of a protein by western blot......

So far I'm still trying to find the optimal concentraion of antibody and protein sample, but I found something like BSA removal kits from varios manufacturers, where BSA is bound specifically and so depleted. E.g:

http://www.millipore.com/catalogue.nsf/doc...74?open&lang=de

offers such a thing. As I said, I haven't tried it myself yet, but maybe it's something for you.... And if it works well, you could inform me...

Mike