一种通过化学发光高灵敏度检测超氧化物歧化酶活性的新技术。
A new technique for highly sensitive detection of superoxide dismutase activity by chemiluminescence.
作者信息
Corbisier P, Houbion A, Remacle J
机构信息
Laboratoire de Biochimie Cellulaire Facultés Universitaires, Namur, Belgium.
出版信息
Anal Biochem. 1987 Jul;164(1):240-7. doi: 10.1016/0003-2697(87)90392-7.
A stable enzymatic free radical generation system has been developed which allows a precise production of 02-. and its detection by chemiluminescence between 2 pmol and 8 nmol. This test has been used for assaying superoxide dismutase (SOD) by inhibition of the chemiluminescence (CL) signal. No inhibition was observed with catalase, which excludes the participation of H2O2 in lucigenin CL. N,N-Diethyldithiocarbamate gives 100% inhibition of SOD activity either from a purified enzymatic preparation or from biological samples, which confirms the specificity of the CL assay. SOD assay can be performed either on a purified enzymatic preparation or on biological materials such as cultured cells.
已开发出一种稳定的酶促自由基生成系统，该系统能够精确产生超氧阴离子（O₂⁻），并通过化学发光在2皮摩尔至8纳摩尔之间对其进行检测。该测试已用于通过抑制化学发光（CL）信号来测定超氧化物歧化酶（SOD）。过氧化氢酶未观察到抑制作用，这排除了过氧化氢（H₂O₂）参与光泽精化学发光的可能性。N，N - 二乙基二硫代氨基甲酸盐对纯化的酶制剂或生物样品中的SOD活性均有100%的抑制作用，这证实了CL测定的特异性。SOD测定既可以在纯化的酶制剂上进行，也可以在生物材料（如培养细胞）上进行。
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