western blot - problem regrding western blotting (Aug/08/2009 )
I am facing a problem in my western blot. I have tried so many time, but after adding the substrate buffer and waiting for 5-15 minutes, let alone to see my band the membrane color remains white even if it stayed overnight or shaked (which is not the normal case). my collegue used the same protein isolation method, same solutions and materials, same protocol and after 2 minutes the bands were there (we are working on different proteins). what do u think is the problem?
-dawitgirma1@yahoo.com-
dawitgirma1@yahoo.com on Aug 8 2009, 12:12 AM said:
I am facing a problem in my western blot. I have tried so many time, but after adding the substrate buffer and waiting for 5-15 minutes, let alone to see my band the membrane color remains white even if it stayed overnight or shaked (which is not the normal case). my collegue used the same protein isolation method, same solutions and materials, same protocol and after 2 minutes the bands were there (we are working on different proteins). what do u think is the problem?
Your antibody isn't working... if everything else is the same.
-bob1-