Glycoprotein detection in nitrocellulose transfers of electrophoretically separated protein mixtures using concanavalin A and peroxidase: Application to arenavirus and flavivirus proteins

✍ Scribed by J.C.S. Clegg

• Book ID: 102984603
• Publisher: Elsevier Science
• Year: 1982
• Tongue: English
• Weight: 810 KB
• Volume: 127
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(82)90192-0

✦ Synopsis

Glycoproteins in nitrocellulose transfers of electrophoretically separated mixtures of cellular and viral proteins are rapidly and sensitively located by sequential incubation with the lectin concanavalin A and the enzymatically active glycoprotein horseradish peroxidase. The bound enzyme is located by incubation with a substrate which is converted to a highly insoluble colored product. The specificity of the method is demonstrated by the abolition of concanavalin A binding in the presence of o-methyl mannoside. The method is capable of detecting as little as 60 ng of a purified model glycoprotein after electrophoresis. It has been applied to the analysis of the glycoproteins of purified Lassa virus and of the virus-specific glycoproteins in Japanese encephalitis virus-infected cells.