Increasing Separation of Bands on SDS PAGE gel - Increasing Separation of Bands on SDS PAGE gel (Mar/02/2010 )
Does anybody have any suggestions as to how I could increase the separation of two bands running ~50 and ~52kDa on a SDS PAGE gel? These bands are partially glycosylated states of a receptor subunit. At the moment the two bands partially merge together. I have run the samples on a 7.5% and on a 10% acrylamide gel but with the same result.
Have you tried 12% or 15% gels? Bands will be higher but may separate. Also I think Tris-Tricine PAGE can give you bit better resolution.
have u tried running at a very slow speed using a high % gel for maybe 2-3hrs? Cos....50 & 52kda is too close.