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[一维毛细管对流免疫扩散法定量测定α1-抗胰蛋白酶]

[Micro-quantitative determination of alpha1-antitrypsin by unidimensional capillary counter-immunodiffusion].

作者信息

Repin V S, Balabanov Iu V

出版信息

Biull Eksp Biol Med. 1977 Apr;83(4):506-8.

PMID:403982
Abstract

The immunoprecipitation reaction for the determination of microamounts of alpha1-antitrypsin was conducted in the capillaries filled with a microvolume of a 1% agarose gel (the gel length was 7--8 mm, and the diameter--0.6 mm). Microamounts of the antigen and antiserum were applied to the surface of the opposite ends of the gel. Precision control of these manipulations was carried out by means of an ocular micrometer of a binocular lens. The amount of the formed immunoprecipitate in gel was calculated by the summated optic density after staining with amido black recorded by the method of double-wave microspectrophotometry. Sensitivity of the method was 0.3--1.0 ng of alpha1-antitrypsin. The linear plot between the amount of alpha1-antitrypsin and the amount of the precipitate formed was found within the range of 1--25 ng of protein. The error was about 10%.

摘要

用于测定微量α1-抗胰蛋白酶的免疫沉淀反应在充满微量1%琼脂糖凝胶的毛细管中进行(凝胶长度为7 - 8毫米,直径为0.6毫米)。将微量抗原和抗血清施加到凝胶相对两端的表面。这些操作的精确控制借助双目镜的目镜测微计进行。凝胶中形成的免疫沉淀物的量通过双波显微分光光度法记录的氨基黑染色后的总光密度来计算。该方法的灵敏度为0.3 - 1.0纳克α1-抗胰蛋白酶。在1 - 25纳克蛋白质范围内发现α1-抗胰蛋白酶的量与形成的沉淀物的量之间呈线性关系。误差约为10%。

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