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BSA contamination in mass spec - (Feb/03/2010 )

Hi! I have a cell line from which I did a membrane extract preparation, and then an immunoprecipitation. Ran the gel, sent it to mass spec and they can only identify BSA. How can I get rid of BSA? Should I wash the cells, use another kind of serum or just use those BSA removal kits? ( they always state that removal of bovine albumin is not optimal... how not optimal??)

-12345678-

12345678 on Feb 3 2010, 02:13 PM said:

Hi! I have a cell line from which I did a membrane extract preparation, and then an immunoprecipitation. Ran the gel, sent it to mass spec and they can only identify BSA. How can I get rid of BSA? Should I wash the cells, use another kind of serum or just use those BSA removal kits?

try washing before trying the removal kits.
( they always state that removal of bovine albumin is not optimal... how not optimal??)

most removal kits will also remove other proteins along with bsa (they use a dye which binds bsa preferentially but not exclusively). if your protein of interest is in low abundance then it may become undetectable if you use the kit.

beckman-coulter is marketing a kit which uses igy to bind bsa. it appears to be specific but it is probably expensive. i would only use it as a last resort.

-mdfenko-

Thank you!! i ll look for the kit and see if it is specific for bovine serum albumin

-12345678-

I am a lab technician working for Protea Biosciences Inc. and the ProtePrep Albumin Depletion Sample Prep kit would be my suggestion for the problem you're having with BSA. This kit offers highly efficient and reproducible binding of approximately 95-99% of BSA from samples without removing any of the lower abundance proteins from the sample. The new affinity-based system allows for removal of albumin more efficiently than anti-body based methods and sample prep times of less than 30 minutes.

-proteaGreg-