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通过透析、凝胶过滤和离子交换色谱联用制备异硫氰酸荧光素标记的γ-球蛋白

PREPARATION OF FLUORESCEIN ISOTHIOCYANATE-LABELED GAMMA-GLOBULIN BY DIALYSIS, GEL FILTRATION, AND IONEXCHANGE CHROMATOGRAPHY IN COMBINATION.

作者信息

DEDMON R E, HOLMES A W, DEINHARDT F

出版信息

J Bacteriol. 1965 Mar;89(3):734-9. doi: 10.1128/jb.89.3.734-739.1965.

DOI:10.1128/jb.89.3.734-739.1965
PMID:14273654
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC277530/
Abstract

Dedmon, Robert E. (Presbyterian-St. Luke's Hospital, Chicago, Ill.), Albert W. Holmes, and Friedrich Deinhardt. Preparation of fluorescein isothiocyanate-labeled gamma-globulin by dialysis, gel filtration, and ion-exchange chromatography in combination. J. Bacteriol. 89:734-739. 1965.-Antiviral immune gamma-globulins isolated from rabbit and guinea pig sera were labeled through dialysis membranes with fluorescein isothiocyanate and purified in several ways to eliminate nonspecific staining. Gel filtration of the conjugate with Sephadex G-25 coarse beads followed by column fractionation with diethylaminoethyl-Sephadex yielded consistently highly specific staining materials. Fluorescein-protein ratios varied between 1.0 and 4.0. This technique has proved to be simple and reliable, and is less time-consuming than previous techniques.

摘要

戴德蒙，罗伯特·E.（伊利诺伊州芝加哥长老会-圣卢克医院），阿尔伯特·W. 霍姆斯，以及弗里德里希·戴恩哈特。通过透析、凝胶过滤和离子交换色谱法联合制备异硫氰酸荧光素标记的γ球蛋白。《细菌学杂志》89:734 - 739。1965年。 - 从兔和豚鼠血清中分离出的抗病毒免疫γ球蛋白通过透析膜用异硫氰酸荧光素进行标记，并通过多种方式进行纯化以消除非特异性染色。用葡聚糖凝胶G - 25粗珠对缀合物进行凝胶过滤，随后用二乙氨基乙基 - 葡聚糖进行柱分级分离，始终能得到高度特异性的染色材料。荧光素 - 蛋白质比率在1.0至4.0之间变化。该技术已被证明简单可靠，且比以前的技术耗时更少。

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