samf45 at attbi.com
Fri Nov 22 01:16:18 EST 2002
Borate interacts with the carbohydrate moiety of glycoproteins while TCA can modify several amino acid side chains in the protein portion. The resulting charge heterogeneity is the cause of the smearing. The solution is to not use borate buffer and not to precipitate with TCA.
suma at iastate.edu wrote:
> I do not have much experience working with proteins. Currently, I am working on cold hardiness studies in Rhododendrons. I am looking for a specific 'Dehydrin' and in SDS-PAGE gels, I see smearing at the top in cold acclimated and non-acclimated samples of one particular species and I do not observe this pattern in case of other species. I am using borate buffer for extraction followed by TCA precipitation. I use exactly same protocol for all the extractions. I think this smearing could be attributed to the carbohydrates and I assume they might not necessarily be bound to the proteins and are not migrating along. They just cause smears!!! and I do not think that there is degradation involved either because, the bands are very sharp. I would definitely appreciate if anyone could help me in this regard.
> Thank you very much!
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