Hi all,
I'm wondering if anyone here have tested anion exchange chromatography for protein fractionation before trypsin digestion instead of using 1Dgel.
I have came across protein fractionation protocol using C18, C4 columns before trypsin digestion, but my sample have SDS, so I'm wondering if ion exchange can help solve the problem.
thanks!
anion exchange chromatography for protein fractionation
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- zougman
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Dear cavaliersun2003,
It is not feasible to perform any type of ion-exchange fractionation on an SDS-solubilized protein sample. You can, however, perform the true size exclusion chromatography (SEC) in this case as was practically demonstrated by Wisniewski et al. http://www.ncbi.nlm.nih.gov/pubmed/20415495
Hope this helps,
AZ
It is not feasible to perform any type of ion-exchange fractionation on an SDS-solubilized protein sample. You can, however, perform the true size exclusion chromatography (SEC) in this case as was practically demonstrated by Wisniewski et al. http://www.ncbi.nlm.nih.gov/pubmed/20415495
Hope this helps,
AZ
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HILIC gets rid of SDS and other surfactants from proteins and peptides quite nicely. What are your proteins? Will they tolerate a predominantly organic solvent?
An alternative is to try anion-exchange with a lot of organic solvent present, although not as much as is necessary for retention in pure HILIC. Say, 40%.
An alternative is to try anion-exchange with a lot of organic solvent present, although not as much as is necessary for retention in pure HILIC. Say, 40%.
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