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Isolating Proteins with SDS Gel Electrophoresis

Isolating Proteins with SDS Gel Electrophoresis - SDS-PAGE Gel Electrophoresis Forum

Isolating Proteins with SDS Gel Electrophoresis - SDS-PAGE Forum and Gel Electrophoresis Forum. Discuss the running of agarose gels, sds-page gels, and other gels including sequencing, gradient or tricine.


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Old 02-22-2011, 05:11 PM
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Question Isolating Proteins with SDS Gel Electrophoresis



Hi, newbie with a question here...

I ran cell lysates under non-reducing conditions and noticed by Western Blot that my protein of interest might be in a disulfide linked complex with other proteins (ran at a high molecular weight). I am interested in finding out what these other proteins might be (I have some guesses). I don't think my PI will pay for mass spec quite yet, so I want to cut out the band(s) I see under non-reducing conditions and re-run them under reducing conditions and then silver stain to see how many proteins might be in the complex. However, I am worried that I won't exactly know what portion of the non-reducing gel to cut out and re-run. Could I silver stain the non-reducing gel before cutting the bands and running under reducing conditions or will that alter the proteins and make re-separation impossible? Any suggestions on how I can identify the proteins in the complex without mass spec is appreciated!

Thanks...
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disulfide , electrophoresis , gel , isolating , protein isolation , proteins , sds , silver stain , western blot


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