On the electrotransfer of polypeptides from gels to nitrocellulose membranes

✍ Scribed by William Lin; Harumi Kasamatsu

• Book ID: 102984808
• Publisher: Elsevier Science
• Year: 1983
• Tongue: English
• Weight: 888 KB
• Volume: 128
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(83)90379-2

✦ Synopsis

The conditions which atfect the elution of polypeptides from polyacrylamide gels by electrophoresis and polypeptide-nitrocellulose interactions have been studied. The rate of elution of polypeptides from a 15% sodium dodecyl sulfate-polyacrylamide gel is dependent on the molecular weight of the individual polypeptides, which is in agreement with the results of W. N. Bumette (Anal. Biochem. 112, 195 (1981)). We also observed that current density affects the rate of elution. Polypeptides smaller than 20,000 daltons pass through pores of 0.45 pm, but not through the pores of 0. l-pm nitrocellulose membranes during electrophoresis. The nonionic detergent NP-40 inhibits the binding of polypeptides to nitrocellulose and removes prebound polypeptides from the membranes. Amido black and Coomassie blue staining and destaining processes do not remove the bound polypeptides from the membranes, but may affect the antigenicity of polypeptides. Polypeptides immobilized on nitrocellulose can be stored at -70°C for future use.