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[醋酸纤维素膜免疫等速电泳]

[Immunoisotachophoresis on cellulose acetate film].

作者信息

Abelev G I

出版信息

Biull Eksp Biol Med. 1978;86(7):110-3.

PMID:678640
Abstract

Acetate-cellulose strips of "Cellogel" type have been shown to be a suitable maintenance medium for performance of isotachophoresis. For immuno-isotachophoresis antigen (from 0.5 to 20 microliter) is applied to a strip of acetate-cellulose film. 1--2 microliter of ampholine solution is placed in front of the antigen zone. All the components present on the strip are made in 0.06 M tris-HCl buffer (pH 6.7), and 0.012 M tris-glycine (pH 8.3) is used as an electrode buffer. Electrophoresis produces migrating Kolraush boundary, which at first is the area of antigen concentration into a narrow starting zone, and then of antigens separation with ampholites. The antigens separated on a cellogel strip are subject to cross-electrophoresis on a film saturated with the respective antiserum, with formation of precipitation peaks for each individual antigen. The method permits to operate with low antigen concentrations since electrophoresis ensures their preliminary concentration and the width of the zones is independent of the time of separation.

摘要

“Cellogel”型醋酸纤维素条已被证明是进行等速电泳的合适维持介质。对于免疫等速电泳,将抗原(0.5至20微升)施加到醋酸纤维素薄膜条上。在抗原区前方放置1 - 2微升两性电解质溶液。条带上存在的所有成分均用0.06 M三羟甲基氨基甲烷 - 盐酸缓冲液(pH 6.7)配制,并用0.012 M三羟甲基氨基甲烷 - 甘氨酸(pH 8.3)作为电极缓冲液。电泳产生迁移的科尔劳施边界,起初这是抗原浓缩到狭窄起始区的区域,然后是抗原与两性电解质分离的区域。在醋酸纤维素条上分离的抗原在用相应抗血清饱和的薄膜上进行交叉电泳,每种单独的抗原都会形成沉淀峰。该方法允许在低抗原浓度下操作,因为电泳可确保其初步浓缩,且区带宽度与分离时间无关。

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