title: Detection of Proteins on Blot Membranes.
abstract: Staining of blot membranes enables the visualization of bound proteins. Proteins are usually transferred to blot membranes by electroblotting, by direct spotting of protein solutions, or by contact blots. Staining allows the efficiency of transfer to the membrane to be monitored. This unit describes protocols for staining proteins after electroblotting from polyacrylamide gels to blot membranes such as polyvinylidene difluoride (PVDF), nitrocellulose, or nylon membranes. The same methods can be used if proteins are directly spotted, either manually or using robotics. Protocols are included for seven general protein stains (amido black, Coomassie blue, Ponceau S, colloidal gold, colloidal silver, India ink, and MemCode) and three fluorescent protein stains (fluorescamine, IAEDANS, and SYPRO Ruby). Also included is an in-depth discussion of the different blot membrane types and the compatibility of different protein stains with downstream applications, such as immunoblotting or N-terminal Edman sequencing. © 2016 by John Wiley & Sons, Inc.
authors: Center for Systems and Computational Biology, and Molecular and Cellular Oncogenesis Program, The Wistar Institute, Philadelphia, Pennsylvania.
authors: Aaron
authors: A
authors: Goldman
authors: Center for Systems and Computational Biology, and Molecular and Cellular Oncogenesis Program, The Wistar Institute, Philadelphia, Pennsylvania.
authors: Sandra
authors: S
authors: Harper
authors: Center for Systems and Computational Biology, and Molecular and Cellular Oncogenesis Program, The Wistar Institute, Philadelphia, Pennsylvania.
authors: David W
authors: DW
authors: Speicher
mesh_terms: D000180:Acrylic Resins / Q000737:chemistry; D004396:Coloring Agents; D004591:Electrophoresis, Polyacrylamide Gel; D005456:Fluorescent Dyes; D015151:Immunoblotting; D008567:Membranes, Artificial; D011506:Proteins / Q000737:chemistry*; D012394:Rosaniline Dyes / Q000737:chemistry; D013194:Staining and Labeling / Q000379:methods*
chemical_list: D000180:Acrylic Resins; D004396:Coloring Agents; D005456:Fluorescent Dyes; D008567:Membranes, Artificial; D011506:Proteins; D012394:Rosaniline Dyes; C016680:polyacrylamide gels; C048139:Coomassie blue
keywords: electroblots; electrotransfer efficiency; membrane stain; protein stain