iodination and immunoprecipitation

Frederick Garbrecht fgarbrec at POST.ITS.MCW.EDU
Thu Feb 18 18:08:27 EST 1993


I would be curious about individuals protocols for radiolabeling cell
surface proteins for immunoprecipitation.  I use a lactoperoxidase based
iodination method using 1-2 mCi 125-I for 50 million cells, to detect
surface proteins with an abundance of about 30-50000 copies per cell.  I
get weak bands (gels take >7 days in the -70), and I suspect that more
iodine is needed.  Does anyone have any simple rules of thumb?  I have
seen protocols that call for as much as 1-2 mCi for each 10 million cells
labeled, but, unfortunately I am not that rich.  Any hints, suggestions?
Thanks
F Garbrecht
Medical College of Wisconsin
fgarbrec at post.its.mcw.edu




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