Preparative elution of proteins blotted to Immobilon membranes

✍ Scribed by Boguslaw Szewczyk; Donald F. Summers

• Book ID: 102984985
• Publisher: Elsevier Science
• Year: 1988
• Tongue: English
• Weight: 909 KB
• Volume: 168
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(88)90008-5

✦ Synopsis

Conditions for the preparative elution of proteins from Immobilon membranes after transfer of proteins to this matrix from sodium dodecyl sulfate-polyacrylamide gels have been established. Proteins were completely eluted from the membrane at room temperature by short incubation in 50 mM Tris-HCl, pH 9.0, containing 2% SDS and 1% Triton X-100. Good protein recoveries were also obtained in the same buffer containing 1% Triton X-100 only. The efficiency of elution was practically independent of the molecular weight of proteins, the method allowed for the precise excision of protein bands, and the proteins eluted from the matrix were not degraded. In some cases it was possible to recover enzymatic activity of the eluted proteins.