title: Analysis of Electroblotted Proteins by Mass Spectrometry.
abstract: Identification of proteins by mass spectrometry is crucial for better understanding of many biological, biochemical, and biomedical processes. Here we describe two methods for the identification of electroblotted proteins by on-membrane digestion prior to analysis by mass spectrometry. These on-membrane methods take approximately half the time of in-gel digestion and provide better digestion efficiency, due to the better accessibility of the protease to the proteins adsorbed onto the nitrocellulose, and better protein sequence coverage, especially for membrane proteins where large and hydrophobic peptides are commonly present.
authors.affiliation: Department of Biochemistry and Molecular Pharmacology and Kimmel Center for Biology and Medicine at the Skirball Institute, New York University School of Medicine, 540 First Ave., New York, NY, 10016, USA.
authors.forename: Jose L
authors.initials: JL
authors.lastname: Luque-Garcia
authors.forename: Thomas A
authors.initials: TA
authors.lastname: Neubert
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D003101:Collodion / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D006801:Humans; D008565:Membrane Proteins / Q000032:analysis; D008567:Membranes, Artificial; D008969:Molecular Sequence Data; D010455:Peptides / Q000032:analysis; D011506:Proteins / Q000032:analysis*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
chemical_list: D008565:Membrane Proteins; D008567:Membranes, Artificial; D010455:Peptides; D011506:Proteins; D003101:Collodion