Recovery of SDS-proteins from polyacrylamide gels by electrophoresis into hydroxylapatite

✍ Scribed by Barry R. Ziola; Douglas G. Scraba

• Publisher: Elsevier Science
• Year: 1976
• Tongue: English
• Weight: 394 KB
• Volume: 72
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(76)90543-1

✦ Synopsis

A method is described which permits the purification of proteins in quantities necessary for physicochemical characterization. The protein to be purified is separated from contaminants by electrophoresis in a number of sodium dodecyl sulfate (SDS)-containing polyacrylamide disc gels. The region of each gel which contains the desired protein band is then cut out and the slices are stacked one above another in a tube. After the protein is electrophoresed from the gel slices into a bed of hydroxylapatite, it is eluted from the hydroxylapatite with 0.5 M sodium phosphate (pH 6.4) containing 0.1% SDS and 1 mM dithiothreitol. Protein recovery is greater than 90%.

Methods

Materials, Lactic dehydrogenase (LDH) and a-chtgn A were obtained from Sigma Chemical Co., carboxypeptidase A (CBP A) and a-chymo-366