Efficient elution of purified proteins from polyvinylidene difluoride membranes (immobilon) after transfer from SDS-PAGE and their use as immunogenes
Following separation of proteins by SDS-PAGE, they are electroblotted onto polyvinylidene difluoride membranes (Immobilon). Protein bands of interest are excised, and the proteins are eluted from the membrane with detergent-containing buffers at pH 9.5. The method routinely yields recovery of 70–90%, and this is independent of protein molecular weight.
Index EntriesPolyvinylidine difluoride protein blotting immunogens Immobilon SDS-PAGE thioredoxin renaturation
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- 3.Hager, D. A. and Burgess, R. R. (1980) Elution of proteins from sodium dodecyl sulfate-polyacrylamide gels, removal of sodium dodecyl sulfate, and renaturation of enzymatic activity: Results with sigma subunit ofEscherichia coli RNA polymerase, wheat germ DNA topoisomerase, and other enzymes.Anal. Biochem. 109, 76–86.PubMedCrossRefGoogle Scholar
- 10.Manson, M. (ed.) (1992)Methods in Molecular Biology, vol. 10: Immunochemical Protocols, Humana, Totowa, NJ.Google Scholar