difference between streaking and smearing of proteins
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streaking of protein lysates that come from mouse spleen tissue on an SDS-PAGE gel (hand cast, not store bought). In this too much protien is a problem so I tried several dilutions (7 ug is the lowest I loaded) and still no dice. It was heard that too much salt is a problem, but we don't know how much is too much? My RIPA buffer that We've used to lyse in is 150mM NaCl. This seems pretty standard, so I'm not sure if this is it. some nasty background on one of the antibodies I use on these samples, and the other one I use is generally clean.
The high concentration of protein blocks the pores of the matrix, making it difficult for proteins to find a path into the gel. the result is a smearing of the bands, as proteins bleed into the gel slowly, instead of entering as a compact band.In general, limit each lane to <20ug of total protein for best results.
The high concentration of protein blocks the pores of the matrix, making it difficult for proteins to find a path into the gel. the result is a smearing of the bands, as proteins bleed into the gel slowly, instead of entering as a compact band.In general, limit each lane to <20ug of total protein for best results.
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