Preparative high-yield electroelution of proteins after separation by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and its application to analysis of amino acid sequences and to raise antibodies

Toshitaka Ohhashi, Chie Moritani, Hiromi Andoh, Sachiko Satoh, Shinji Ohmori, Friedrich Lottspeich, Mikiko Ikeda

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

A method for the preparative high-yield electroelution of proteins from sodium dodecyl sulphate (SDS) polyacrylamide gel strips was established. The method consisted of SDS-polyacrylamide gel electrophoresis, detection of proteins with sodium acetate and electrophoretic elution at 200 V for 3 h by utilizing a horizontal flat-bed gel electrophoresis apparatus. Standard proteins with molecular masses of 14-66 kilodalton (cytochrome c, aldolase, ovalbumin and bovine serum albumin) were recovered with an average yield of 73.6 ± 2.3%. A membrane-bound protein, rat skeletal muscle Ca2+-ATPase (100 kilodalton) was also well recovered (over 60%). This method was applicable to the purification of proteins required for N-terminal amino acid sequencing and to raise antibodies.

Original languageEnglish
Pages (from-to)153-159
Number of pages7
JournalJournal of Chromatography A
Volume585
Issue number1
DOIs
Publication statusPublished - Oct 25 1991

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Organic Chemistry

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