title: Microwave-assisted protein staining, destaining, and in-gel/in-solution digestion of proteins.
abstract: Rapid evolution of state-of-the-art proteomic analyses has encompassed development of high-throughput analytical instrumentation and bioinformatic tools. However, recently, there has been a particular emphasis on increasing the throughput of sample preparation, which has become one of the rate-limiting steps in protein characterization workflows. Researchers have been investigating alternative methods to conventional convection oven incubations to try and reduce sample preparation time for protein characterization. Several protocols have appeared in the literature, which employ microwave irradiation as a tool for the preparation of biological samples for subsequent characterization by a variety of analytical techniques. In this chapter, techniques for microwave-assisted protein staining, destaining, and digestion are described. In general, the application of microwave-assisted technologies resulted in the drastic reduction of overall sample preparation time, though discrepancies in the reproducibility of several published digestion protocols still remain to be clarified.
authors.affiliation: Genentech, Inc., South San Francisco, CA, USA.
authors.forename: Jennie R
authors.initials: JR
authors.lastname: Lill
authors.forename: Victor J
authors.initials: VJ
authors.lastname: Nesatyy
mesh_terms: D000818:Animals; D002417:Cattle; D004396:Coloring Agents / Q000737:chemistry; D016898:Interferon-alpha / Q000737:chemistry / Q000302:isolation & purification; D007700:Kinetics; D008872:Microwaves; D009942:Organometallic Compounds / Q000737:chemistry; D010449:Peptide Mapping / Q000379:methods; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification; D059748:Proteolysis; D012394:Rosaniline Dyes / Q000737:chemistry; D012710:Serum Albumin, Bovine / Q000737:chemistry* / Q000302:isolation & purification; D016622:Silver Staining / Q000379:methods; D013194:Staining and Labeling / Q000379:methods; D014357:Trypsin / Q000737:chemistry
chemical_list: D004396:Coloring Agents; D016898:Interferon-alpha; D009942:Organometallic Compounds; D011506:Proteins; D012394:Rosaniline Dyes; C515601:Sypro Ruby; D012710:Serum Albumin, Bovine; C048139:Coomassie blue; D014357:Trypsin