western blot problems

v-liu2 at northwestern.edu v-liu2 at northwestern.edu
Thu Apr 22 16:36:12 EST 2004


I have been having all kinds of problems with western blot.  Recently, I have run into a very frustrating problem.  After 2 re-blots, I did GAPDH for loading control, and there was nothing!  However, the positive control lysate was fine, but my samples had no GAPDH at all!  The fresh membrane had good banding pattern, but after the first re-blot, the bands were really faint, and after the 2nd re-blot, not even GAPDH showed up!  I am really frustrated with it.  Does somebody have similar experience?  Could anybody help me out?  Is it a problem with the protein lysate?  

Thank you!



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