A simplified chloral hydrate electrophoresis system for analysis of biological membranes

✍ Scribed by Byron Ballou; Oliver Smithies

• Publisher: Elsevier Science
• Year: 1977
• Tongue: English
• Weight: 586 KB
• Volume: 80
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(77)90686-8

✦ Synopsis

A chloral hydrate-containing tube-gel electrophoresis system previously described by Ballou, Sundharadas. and Bach [( 1974) Science 185, 5311 permitted charge and size separation of membrane proteins, but was difficult to use and inapplicable to slab gels. A greatly simplified photopolymerized chloral hydratepolyacrylamide gel electrophoresis system is presented here for use with tube or slab containers. An aluminum lactate buffer system provides good solubilization and resolution. Its advantages over the previously described system are simplicity and multiple sample capacity.

Electrophoresis in polyacrylamide gels containing concentrated chloral hydrate solutions was shown by Ballou et al. ( 1) to be an effective means of analyzing proteins from biological membranes; however. the system originally described used highly purified reagents and required considerable care in polymerizing the acrylamide gel matrix. We describe here a simpler and more flexible chloral hydrate-slab gel electrophoresis system which needs little time and labor to yield equally good resolution. The method permits the use of technical grade reagents throughout, except for the buffer, and allows direct comparison of many samples run on the same gel. It should be particularly suitable in initial screening for allelic variants of membrane systems.

Methods

Apparatus.