TY - JOUR T1 - A modified FASP protocol for high-throughput preparation of protein samples for mass spectrometry JF - bioRxiv DO - 10.1101/084533 SP - 084533 AU - Jeremy Potriquet AU - Marut Laohaviroj AU - Jeffery Bethony AU - Jason Mulvenna Y1 - 2016/01/01 UR - http://biorxiv.org/content/early/2016/10/31/084533.abstract N2 - To facilitate high-throughput proteomic analyses we have developed a modified FASP protocol which improves the rate at which protein samples can be processed prior to mass spectrometry. Adapting the original FASP protocol to a 96-well format necessitates extended spin times for buffer exchange due to the low centrifugation speeds tolerated by these devices. However, by using 96-well plates with a more robust polyethersulfone molecular weight cutoff membrane, instead of the cellulose membranes typically used in these devices, we could use isopropanol as a wetting agent, decreasing spin times required for buffer exchange from an hour to 30 minutes. In a typical work flow used in our laboratory this equates to a reduction of 3 hours per plate. To test whether our modified protocol produced similar results to FASP and other FASP-like protocols we compared the performance of our modified protocol to the original FASP and the more recently described eFASP and MStern-blot. We show that all FASP-like methods, including our modified protocol, display similar performance in terms of proteins identified and reproducibility. Our results show that our modified FASP protocol is an efficient method for the high-throughput processing of protein samples for mass spectral analysis.AbbreviationsABCAmmonium bicarbonateACNAcetonitrileTFATrifluoro acetic acidFAFormic acidTEABTriethylammonium bicarbonateDCADeoxycholic acidDTTDithiothreitolIAAIodoacetamide ER -