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Pulse-Chase Protocols needed

Ronald Berry Berry at biochem.unp.ac.za
Tue Aug 31 04:37:11 EST 1993


Hello out there,
Our lab is about to embark on a pulse-chase protein labelling experiment. 
We want to pulse some animal cell cultures with 35S-Methionine, then harvest 
at various time intervals. The protein of interest is to be immuno-
precipitated, followed by SDS PAGE and autoradiography to follow its 
turnover. The protein inquestion exists in 2 forms -22kD and 26kD, and we 
need to resolve this pair.
My problem is that I cannot find a step-by-step protocol for this type of 
work. It is either so routine that the fokelore has become absorbed into 
general texts, or so specialised that noone wants to try (or am I being too 
cynical??!).
Does anybody have a Child's Guide, especially to the radiolabelling/
autoradiography side ot this type of work?
Any help wold be gratefully appreciated.
Thanks
Ron Berry
Chief Technician 
Department of Biochemistry
University of Natal
Pietermaritzburg
Natal
South Africa



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