In article <[email protected]>, [email protected] says... > > Hello, I am able to enrich and immunoprecipitate a very large glycoprotein > from human epithelial cells. I suspect that it undergoes ectodomain > shedding as I can detect it in the media. To analyze trafficking I > attempted to use S35 labeling which repeatedly failed with only background > bands showing up on the radiogram. Various pre-clearing steps to reduce > background did not help. Is it possible that S35 pulse chase is simply not > suitable to study proteins expressed at very low levels? Does Cycloheximide > chase substitute for S35 pulse chase? > > Thank you.
How many Cys and Met does your protein have (in its processed form)? Since it is a glycoprotein, you could try radioactively labelled sugars, but those are finnicky to use. _______________________________________________ Methods mailing list [email protected] http://www.bio.net/biomail/listinfo/methods
