I am doing silver staining since my protein concentration is low. However, the gel became dark and the protein bands appear as negative band (even the bands which are not present in coomassie staining). I made a new develop solution, this time I could see some fade brown/black bands instead of negative bands but it took very long time (> 20 minutes) to develop and lots of background. I wonder why my bands cannot appear clearly as shown in figures about silver staining
Quynh Pham
@ Dirk Schmidt: I do not use commercial silver staining kit but I found the reason. The first trial, I used exhausted thiosulfate and the second time, I used recycled running buffer. I changed all the reagent and my staining looks well now.
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