A microtiter-plate assay of nitric oxide synthase activity

被引：15

作者：

Dawson, J

Knowles, RG

机构：

[1] Glaxo-Wellcome Research and Development, 5G120, Gunnels Wood Road, Stevenage, Herts

来源：

MOLECULAR BIOTECHNOLOGY | 1999年 / 12卷 / 03期

关键词：

nitric oxide synthases; oxyhemoglobin; microtiter-plate; kinetic;

D O I：

10.1385/MB:12:3:275

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

We describe here a microtiter-plate assay for measuring nitric oxide synthase (NOS) activity by utilizing the spectral shift in optical absorbence between the wavelengths 405 and 420 nm on conversion of oxyhemoglobin to methemoglobin by nitric oxide (NO). This is a high-throughput assay permitting 96 or 384 Simultaneous kinetic measurements and is ideal for the study of NOS inhibitors and their time dependence. It is also possible to measure enzyme rates under different conditions simultaneously for the study of the cofactor and substrate dependence of NOS preparations. The assay requires approximately 10 pmol/min of NOS activity to achieve a 1 moD/min rate.

引用

页码：275 / 279

页数：5

共 12 条

[1]

[Anonymous], [No title captured]

[2]

Charles IG, 1996, METHOD ENZYMOL, V268, P449

[3]

FEELISCH M, 1987, EUR J PHARMACOL, V139, P9

[4] CYTOKINE-ACTIVATED ENDOTHELIAL-CELLS EXPRESS AN ISOTYPE OF NITRIC-OXIDE SYNTHASE WHICH IS TETRAHYDROBIOPTERIN-DEPENDENT, CALMODULIN-INDEPENDENT AND INHIBITED BY ARGININE ANALOGS WITH A RANK-ORDER OF POTENCY CHARACTERISTIC OF ACTIVATED MACROPHAGES [J].