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Radioactive Pulse-Chase Analysis and Immunoprecipitation
#MMPMID28480315
Magadán JG
Bio Protoc
2014[Apr]; 4
(8
): ? PMID28480315
show ga
Labeling of newly-synthesized polypeptides with radioactive amino acids followed
by immunoprecipitation allows quantitative analysis of the fate of a given
protein in a time-dependent manner. This biochemical approach is usually used to
study a variety of processes, such as protein folding, co-translational
modifications, intracellular transport, and even its rate of degradation. Here, I
describe step by step a simple technique to both label newly-synthesized
influenza A virus (IAV) hemagglutinin (HA) with [(35)S]-methionine and then
follow its maturation and transport through the secretory pathway by SDS-PAGE and
fluorography (Magadan et al., 2013).