After performing silver staining on the 12% resolving gel containing proteins, the upper part of the gel became dark brown with translucent spots but the lower part of the gel had dark brown spots with a very light background. After repeating the staining procedure on another gel , the background became dark brown in color with no spots. But when the same gel was kept in coomassie blue stain overnight, a few protein spots were seen. I have repeated the experiments on new gels but got same negative results!
I have used G E healthcare plus one silver staining kit for proteins.
Hi Sania!!
there could be many possibilities for getting negative results. I assume you did 2DE. Then, initial protein quantity, the separation in 1st dimension, protein quality as well as your staining procedure could be the reasons. So, initially try to look into specific problem.
Silver stain is very sensitive and can detect upto ng levels of protein.
all the best!!
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