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采用碘甘法用¹²⁵I标记血小板表面蛋白。

Labeling of platelet surface proteins with 125I by the iodogen method.

作者信息

Tuszynski G P, Knight L C, Kornecki E, Srivastava S

出版信息

Anal Biochem. 1983 Apr 1;130(1):166-70. doi: 10.1016/0003-2697(83)90664-4.

Abstract

A procedure for the 125I-iodination of platelet suspensions is described. The procedure utilizes Iodogen, a solid-phase oxidizing agent similar to chloramine-T. Platelets were labeled under a variety of conditions, including in the presence of 0.1% albumin, and showed between 7 and 28% incorporation of 125I. Best labeling results were obtained at low platelet concentrations (3-5 x 10(8) platelets/ml), short reaction times (15 min), and with 2-ml glass vials coated with 100 micrograms of Iodogen. Analysis of the labeled platelet proteins by sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by autoradiography revealed that the same major protein bands were labeled by this procedure as were labeled by the lactoperoxidase procedure. At low platelet concentrations, the Iodogen procedure gives twice the amount of iodine incorporation.

摘要

本文描述了一种血小板悬液的¹²⁵I碘化方法。该方法使用碘甘醚,一种类似于氯胺-T的固相氧化剂。血小板在多种条件下进行标记,包括在0.1%白蛋白存在的情况下,¹²⁵I的掺入率在7%至28%之间。在低血小板浓度(3 - 5×10⁸个血小板/毫升)、短反应时间(15分钟)以及用100微克碘甘醚包被的2毫升玻璃小瓶的条件下,获得了最佳标记结果。通过十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳随后进行放射自显影对标记的血小板蛋白进行分析,结果显示该方法标记的主要蛋白带与乳过氧化物酶方法标记的相同。在低血小板浓度下,碘甘醚方法的碘掺入量是其两倍。

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