Routine identification of proteins from sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gels or polyvinyl difluoride membranes using matrix assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS).
As the resource laboratory for Rockefeller University our emphasis continues to be on methodology development for the routine analysis of low abundance proteins isolated from native sources. In the past ten years, gel electrophoresis of proteins has become the method of choice for the preparation of microgram and submicrogram quantities of protein for primary structural characterization, and over 95% of the samples submitted for protein identification are either in a gel or bound to polyvinyl difluoride membranes (PVDF). As such, we employ multiple microanalytical approaches encompassing Edman sequence degradation, amino acid and matrix assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometric analysis to provide an integrated protein characterization of such samples. Here we describe the two major services we employ when providing protein identification from in-gel or PVDF-bound proteins.
Fernandez
J
J
The Rockefeller University Protein/DNA Technology Center, New York, NY 10021, USA.
Gharahdaghi
F
F
Mische
S M
SM
D000595:Amino Acid Sequence; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D006868:Hydrolysis; D008567:Membranes, Artificial; D008969:Molecular Sequence Data; D010447:Peptide Hydrolases / Q000378:metabolism; D010449:Peptide Mapping / Q000379:methods*; D011145:Polyvinyls; D011506:Proteins / Q000302:isolation & purification*; D012967:Sodium Dodecyl Sulfate; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D005465:Fluorocarbon Polymers
D008567:Membranes, Artificial; D010447:Peptide Hydrolases; D011145:Polyvinyls; D011506:Proteins; D012967:Sodium Dodecyl Sulfate; C024865:polyvinylidene fluoride; D005465:Fluorocarbon Polymers