125I-labeling of platelet proteins with Bolton-Hunter reagent

✍ Scribed by Gregg E. Davies; Jiri Palek

• Book ID: 102984474
• Publisher: Elsevier Science
• Year: 1981
• Tongue: English
• Weight: 859 KB
• Volume: 115
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(81)90021-x

✦ Synopsis

Treatment of washed, intact platelets with Bolton-Hunter reagent is a satisfactory method for '251-labeling of many platelet proteins. Analysis by two dimensional polyacrylamide gel electrophoresis and autoradiography shows that the major platelet cytoskeletal proteins and at least four surface-exposed proteins are labeled. The method allows the identification of these labeled proteins in amounts that are below the limits of detection by Coomassie blue staining. Two granule proteins, thrombospondin and fibrinogen, are slightly labeled. Conditions of labeling do not appear to affect platelet structure or function, as assessed by phase-contrast microscopy, "Cr04'-release, and aggregation in response to thrombin or fibrinogen/adenosine-5'-diphosphate. ' Abbreviations used: Hepes. 4-(2-hydroxyethyl)-lpiperazineethanesulfonic acid; SDS, sodium dodecyl sulfate.