Document Type
Journal Article
Publication Date
1-1-2017
Journal
PLoS One
Volume
12
Issue
7
DOI
10.1371/journal.pone.0175967
Abstract
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 typical work flow used in our laboratory this equates to a reduction of 3 hours per plate, providing processing times similar to FASP for the processing of up to 96 samples 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.
Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 License.
APA Citation
Potriquet, J., Laohaviroj, M., Bethony, J., & Mulvenna, J. (2017). A Modified FASP Protocol for High-throughput Preparation of Protein Samples for Mass Spectrometry.. PLoS One, 12 (7). http://dx.doi.org/10.1371/journal.pone.0175967
Peer Reviewed
1
Open Access
1
Included in
Biomedical Engineering and Bioengineering Commons, Medical Immunology Commons, Medical Microbiology Commons, Tropical Medicine Commons
Comments
Reproduced with permission of PLoS ONE