Metachromasy of peripheral nerve collagen on polyacrylamide gels stained with Coomassie brilliant blue R-250

✍ Scribed by Steven Micko; William W. Schlaepfer

• Publisher: Elsevier Science
• Year: 1978
• Tongue: English
• Weight: 569 KB
• Volume: 88
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(78)90457-8

✦ Synopsis

Endoneurial collagen stains metachromatically with Coomassie brilliant blue R-250 (C.I. 42660) when peripheral nerve proteins are solubilized with urea and SDS and then subjected to SDS-polyacrylamide gel electrophoresis. The metachromasy is reproducible under different fixing and staining conditions, but was exhibited only by Coomassie blue R-250 of the four triphenylmethane dyes tested. A method is presented for measurement of the degree of metachromasy on SDS gels and the detection of collagen in homogenates of whole tissue.

Methods

Peripheral nerve proteins and polypeptide standards. Sciatic, femoral, and axillary nerves and nerve roots were dissected and desheathed as described previously (1). Desheathed nerve segments were solubilized in 8 M urea and 1% SDS by the method of Hoffman and Lasek (5) except