Does anybody has an experience with commercial kits for depletion of the most abundant proteins (IgG, albumin, etc.) for further analysis of low-abundant proteins?
Thank you very much.
you can use hydrophobic interaction chromatography, attached you can find a paper in this topic
Article Depletion of highly abundant proteins in blood plasma by hyd...
The Pierce Top 2 and ABI/GE HSA IgG depletion spin columns have worked well for me in 2D gels. They typically use a protein A/G to pull out IgG and either an anti HSA MAb or old fashioned dye binding column to remove HSA. Pierce has a newer (Top12) column which they say removes the 12 most abundant human serum proteins (!) but I have not tried it.
We've compared the Pierce Top12 with Sigma ProteoPrep (cartridges) as well as Biorads ProteoMiner. The ProteoPrep, at least in our hands, provides the deepest coverage although not by a big difference. Using the ProteoPrep system can do a double depletion but end up with very little material to work with.
The Mars columns are popular but have not tested. But whatever u decide to use will still end up with a fair bit of abundant proteins so my suggestion would be to use a targeted method if wanting to monitor low abundance species
Yes ... you can use the kits to deplete High abundance Sigma ProteoPrep (cartridges). I already used it and it works well. After that its better to make it concentrate
Dear all,
thank you for your answers and suggestions. I appreciate it. Have you tried immunodepletion resins instead of columns and spin cartridges? Do you have any comparison to this (regarding price, time...)? I would like to use one of spin-down column or immunodepletion resin (not LC colums) for further glycomic analysis of PSA.
Apologies for resurrecting the topic, but I came across your question whilst searching for something else and realised I have an answer to contribute - we have extensively characterised MARS6, MARS14 and ProteoPrep20 columns in human serum, and analysed the results by 1D-PAGE, 2D-DIGE and 1D-LC-MS/MS, looking at efficiency of depletion, specificity, reproducibility and depth of proteomic coverage
Link: http://www.ncbi.nlm.nih.gov/pubmed/21548096
We've also used our preferred one (MARS14) to discover serum biomarkers of potential diagnostic and prognostic value in human renal transplantation
Link: http://www.ncbi.nlm.nih.gov/pubmed/23739232
I hope this is helpful, and apologies I was not able to join the discussion sooner!
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