hello all,
i'm working on a Bio-Rad mini 2-D gel system. we just got it in recently and are trying to get our protocol down. we are using Bio-Rad's 2-D protein markers as our guide and visualizing using silver stain (Bio-Rad too). however, we seem to be having a problem resolving the more basic proteins of this marker cocktail (>pH 6.0), while the more acidic proteins are fine. to troubleshoot this, we have done:
1.) increased focusing time to 7 hours at 750 V.
2.) decreased protein load
do you kind people have any other suggestions for me to resolve these more basic proteins better while keeping the same quality of acidic resolution? i would really appreciate it.
Aaron Downs
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