quantification of protein
rogier
stugerNOstSPAM at cellbiology.uni-frankfurt.de.invalid
Tue Oct 5 13:29:35 EST 1999
Hiya,
I'd say do a quantitative western (wetblotting, not semi-dry). make
sure you load equal amounts of protein by doing a lowry assay on your
protein in SDS-PAGE loading buffer (the spot method in Anal Biochem 223
(1994) 329-31 works OK).
Ponceau your blot (and make sure you blot the entire gel, including the
stacking) to make sure equal amounts of protein in all lanes managed to
enter your gel as opposed to staying in the slots or collecting on the
border of stacking/running gel.
We routinely do this on E. coli proteins with less than 10% error.
Good luck,
Rogier
rogier at biogate.com
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