I had solubilized organellar pellet with (1%SDS, 0.1N NaOH) and resolved in gel in presence of 1x SDS sample buffer, but the separation of the proteins does not look that good. I would like to precipitate my proteins and separate from other interfering factors. But now they are in SDS sample buffer.
Shreyasi Asthana
Hi Jeevan
What all your sample buffer contain?I think adding TCA at this point is not a good idea
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Venil N Sumantran
You can dilute 5 times, to get 0.2% SDS and 0.02NNaOH. Then, TCA pptn can be done. But, in my experience, you only get back 60% of the starting protein concentration. Acetone pptn maybe better. If you have enough protein, you could compare both methods! Hope this helps
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