I'm looking for a MS-compatible approach to elute immunoprecipitated proteins prior to proteolysis. Does anyone have experience with using Rapigest (or similar acid-labile surfactants) for this purpose? Ideally, I would like to obtain quantitative recovery - what sort of concentration would be required to achieve this?
Share protocols and ask for sample preparation advice.
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- Phosphoserine Member
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- Angiotensin Member
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We elute our IPs in Laemlli buffer and then clean them up prior to proteolysis using SP3 (http://www.ncbi.nlm.nih.gov/pubmed/25358341). You can use whatever version of Laemlli that you want, with or without glycerol, with our without dye. We also elute in RIPA sometimes. SP3 will get rid of whatever components you use in your elution. Then you can go straight to MS after your digestion without having to clean up the Rapigest.
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