A dot-blot assay for heparin-binding proteins

✍ Scribed by Nobuyoshi Hirose; Michele Krivanek; Richard L. Jackson; Alan D. Cardin

• Publisher: Elsevier Science
• Year: 1986
• Tongue: English
• Weight: 794 KB
• Volume: 156
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(86)90260-5

✦ Synopsis

A method for the detection and quantitation of picomole amounts of heparin-binding proteins is described. Proteins are first spotted on nitrocellulose and then incubated with "51-heparin. Binding of heparin to the proteins is detected by radioautography and quantitated by scanning densitometry;

proteins are quantitated by densitometric analysis of the amido black stained nitrocellulose.

Heparin-binding was time-dependent and sensitive to the presence of metal ions. urea, and detergents (anionic, nonionic, and zwitterionic).

The divalent cations Ca*' and Mg" and the zwitterionic detergent 3-[(3-cholamidopropyl)dimethylammonio]-I-propanesulfonate increased heparin binding whereas NaCI, urea, sodium dodecylsulfate, and La3+ decreased binding. This assay is applicable to the identification and characterization of a variety of heparin-binding pI'OteiIlS.