Dear colleagues,
We've tried to perform Co-immunoprecipitation on human cell line samples.
After the Co-IP, we ran gels, excise individual bands of interest for MALDI-TOF/TOF protein identification.
Nevertheless, many of the samples seemed to match BSA. But some of these bands did not conform to the MW of BSA (e.g. bands at ~35kDA in SDS-PAGE also matched BSA).
We've double checked our MALDI-TOF/TOF sample preparation and workflow. It seemed to be alright (other samples processed together showed trust-worthy match to other proteins).
Have you encountered similar situation before? What would be the source of BSA contamination (Is BSA contamination commonly observed in Co-IP of human cell lines)?
Thank you!
If you are getting BSA contamination from human cell lines that means the beads you are using have been blocked with BSA before. Try using fresh beads and instead of blocking the beads with BSA, do a pre-clearing of your samples with the beads. It should work fine.
If the human cell line was grown in medium containing fetal bovine serum, the contamination with BSA could be from the serum, which has a very high concentration of albumin.
One common source of BSA contamination could be the box that you use to put your gel for staining. Someone might have used that box for blocking their western blot membrane. So its better to use new box and wash gel apparatus properly before use.
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