Hi, I have seen several semi-native SDS PAGE protocols.
I am wondering if the semin native SDS-PAGE can be used to accurately estimate protein molecular weight? Is the added SDS enought to give all the proteins the same charge density (according to some protocols just 0,1% to the running buffer and none to the sample and the gel)?
According to my understanding, adding SDS to the protein and boiling it is the main reason that denatures the protein and allows SDS to surround it and give an overall charge. Adding SDS to only the running buffer may not allow complete denaturation and the protein might run only on the basis of its molecular weight and the isoelectric point. Without SDS, different proteins with similar molecular weights would migrate differently due to differences in mass-charge ratio.
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