Protocol Online logo
Top : New Forum Archives (2009-): : SDS-PAGE and Western Blotting

Methods of concentrating total protein extracts - (May/20/2012 )

How can I concentrate my protein extract which would give me the best final yield?

My question arises from the feint Western blotting band which indicated low protein expression. I've already loaded 30-40ug protein. Should I load more? concentrate my stocks?

-science noob-

I have had the same problem with a very low expressed group of proteins. I use a large gel that I have to make since no one sells them and make a large well, then I add 100 ug of protein and use an biotination secondary antibody and a senstive ECL kit like Pierce's West pico.


i precipitated mine with acetone.. and got concentrated pretty well.. now i see my bands clearly