Biomedical Engineering and Bioengineering Commons^™

A Modified Fasp Protocol For High-Throughput Preparation Of Protein Samples For Mass Spectrometry., Jeremy Potriquet, Marut Laohaviroj, Jeffrey M Bethony, Jason Mulvenna

Microbiology, Immunology, and Tropical Medicine Faculty Publications

o 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 …