Help in SDS PAGE - Molecular Biology (Mar/30/2010 )
I am running a SDS-PAGE for the first time, I have ordered pre cast gels from Biorad and I wanted to know what to use as sample loading buffer and running buffer.
I understand that the Biorad website is pretty hard to navigate but this should (hopefully) provide all the information you need.
In short, for standard SDS-PAGE your running buffer is made up of Tris, SDS and Glycine. Sample buffer has Tris (pH 6.8), Glycerol, SDS, bromophenol blue and either DTT or 2-ME in it for reducing. Recipes for both can be found in the pdf.
Remember that you also might need sliver stain or coomassie blue to see your proteins, or if using antibody detection, membrane, ECL developing reagent, antibodies etc.
Hope it works out for you.
SDS Page is sth. really rudiment and there should be a lot of recipes around in the big thing called internet. But probably just check the lab next door, they have some buffers and can give you the recipe as well.
Remember also that this is Protocol-online, so there are lots of protocols on this site alone.