I want to prepare 5X SDS sample buffer, some protocol has 2% SDS some has 10% SDS, is there any difference between the two?
why different protocols have different percent ,is it depend on the protein that i extract?
5x sample buffer just implies that you will make a dilution of your sample such that the ending buffer concentration is 1x (4 mL of sample + 1 mL of 5x buffer). In your case the final, desired SDS concentration is 2%.
It is my experience that, provided that the sample to be analyzed does not contain too much salt, 1x Lämmli sample buffer (i.e. 2% SDS) works fine even if diluted 1:1 with the sample. In most cases, the difference in detection sensitivity between a sample that is diluted to 50% (1:1, 1 x buffer) vs a sample diluted to 80% (4:1, 5x buffer) is not crucial. Thus, I use 1 x buffer for all samples and don't bother to make 5x buffer.
It depends which one works better in your case, usually if we didn't get the perfect results, we do some minor changes in the protocol to get the perfect results
so that's why different labs have slight variations in protocol.
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