DETERMINATION OF PLATELET-ACTIVATING-FACTOR ACETYLHYDROLASE ACTIVITY BY BLOTTING, BETA-RADIOLUMINESCENCE, AND ULTRAHIGH-SENSITIVITY TELEVISION CAMERA DETECTION

被引:6
作者
AKIYAMA, M
SUGATANI, J
SUZUKI, Y
MIWA, M
机构
[1] UNIV SHIZUOKA,SCH PHARMACEUT SCI,DEPT BIOCHEM,SHIZUOKA 422,JAPAN
[2] KANSAI MED UNIV,DEPT MED CHEM,MORIGUCHI,OSAKA 570,JAPAN
关键词
D O I
10.1006/abio.1994.1181
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
A simple method for detecting the 1-alkyl-2-[H-3]acetyl-sn-glycero-3-phosphocholine ([H-3]acetyl-PAF)-hydrolyzing activity of serum proteins is described. These were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to polyvinylidene difluoride (PVDF) membranes. The assay involves measurement of the beta-radioluminescence of [H-3]acetyl-PAF on the PVDF membranes with an ultrahigh-sensitivity TV camera system. Enzyme activity is detected as a decrease in beta-radioluminescence of [H-3]acetyl-PAF on the blotted membranes, since of [H-3]acetyl-PAF remains on the membrane but the [H-3]acetate released by the enzyme reaction detaches from the membrane. This method has been used to identify PAF acetylhydrolase and phospholipase A(2). (C) 1994 Academic Press, Inc.
引用
收藏
页码:295 / 299
页数:5
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