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Western blot using blood plasma - (Aug/15/2013 )

If I wish to examine phosphoprotein levels in plasma samples using Western blot, do I just use the samples directly for the experiment like protein lysates? Are there any preparations or taboos I should be aware of?

-polar41323-

you may find that you have to deplete albumin to get a good look at all of the phosphoproteins.

-mdfenko-

Will the use of albumin depletion kit do the job? A reference paper warned that there might be non-specific loss which may affect the analysis.

-polar41323-

there are some albumin depletion kits that are very specific for albumin (such as this one from sigma).

-mdfenko-

This is very helpful!! Thank you!!

-polar41323-

If I performed immunoprecipitation before western blot, will it decrease the non-specific signals? if so, can I directly use the plasma for pull-down? The depletion of albumin should be done BEFORE the immunoprecipitation, right?

-polar41323-