S35- met labelling of proteins: URGENT

wetsel_r at wums.wustl.edu wetsel_r at wums.wustl.edu
Thu Jul 30 09:47:24 EST 1992


That's an open question, some additional details would help.  However, we
routinely label cultured cells using the 35S-Trans label from ICN.  We bring up
the volume such that 100 ul equals 1mCi.  We plate our cells in either 24 or 6
well plates at 3-4X10^5 or 1X10^6 cells per well, respectively.  Use met free
 medium with either 0.5 % BSA or 5% dialyzed calf serum.  Depending upon the
cell type, we will add 0.250 - 0.500 uCi per ml of labeling media and incubate
that on the cells anywhere from 2-24 hours, normally 4 hours at 37'C works very well for most continuous cell lines.  I should note, prior to adding the
labeled media to the cells, we will wash the cells 2-3 times in HBSS.  This
works very well in our hands for both adherent and non-adherent cells.  We oftenwill then use the corresponding supernatant and cellular lysate in
immunoprecipitations - I'll assume that is what you're doing?  Post back
if you need more help... best of luck...

David



More information about the Methods mailing list