Prince:Filter-aided Sample Preparation

Brief Explanation

Filter-aided sample preparation (FASP) prepares protein for mass spectrometry. After lysis, proteins are placed on a filter and washed repeated to remove detergent. Detergents creates noise in the mass spectrometer and contaminate the chromatography. After a number of washing steps the proteins are ready for protease digestion. Proteases are enzymes that cut proteins into peptides. We normally use trypsin, which cuts after Lysine and Arginine amino acids. Trypsin digestion normally takes 18 hours.

Yeast FASP

Literature

1. Wisniewski, J. R.; Zougman, A.; Nagaraj, N.; Mann, M., Universal sample preparation method for proteome analysis. Nat. Methods 2009, 6 (5), 359-U60. http://www.nature.com/nmeth/journal/v6/n5/full/nmeth.1322.html