title: Optimization of cell lysis and protein digestion protocols for the analysis of HeLa S3 cells by LC-MS/MS.
abstract: In order to maximize the number of proteins identified from Hela S3 cell lysate we tested various cell lysis, protein precipitation and digestion protocols. First, we compared three different lysis buffers, two mechanical cell disruption methods and two precipitation methods. Then, we tested six different in-solution digestion protocols, three different in-gel digestion protocols and ten different peptide extraction protocols. The result is a proposal for an optimized protocol to prepare the whole cell lysate samples from HeLa S3 cells.
authors: Department of Pathology, Children's Hospital Boston and Harvard Medical School, Boston, MA 02115, USA.
authors: Dominic
authors: D
authors: Winter
authors: Hanno
authors: H
authors: Steen
mesh_terms: D002853:Chromatography, Liquid / Q000379:methods; D006367:HeLa Cells; D006801:Humans; D011506:Proteins / Q000378:metabolism; D059748:Proteolysis; D020543:Proteome / Q000032:analysis*; D040901:Proteomics / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D011506:Proteins; D020543:Proteome