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
Does anybody have a Child's Guide, especially to the radiolabelling/
autoradiography side ot this type of work?
Any help wold be gratefully appreciated.
Department of Biochemistry
University of Natal