文章基本信息

标题：Solid-Phase Conjugation of Ferritin to Fab-Fragments of Immunoglobulin G for Use in Antigen Localization on Thin Sections
本地全文：下载
作者：J. P. Kraehenbuhl ; J. D. Jamieson
期刊名称：Proceedings of the National Academy of Sciences
印刷版ISSN：0027-8424
电子版ISSN：1091-6490
出版年度：1972
卷号：69
期号：7
页码：1771-1775
DOI：10.1073/pnas.69.7.1771
语种：English
出版社：The National Academy of Sciences of the United States of America
摘要：A technique for the solid-phase conjugation of ferritin to monovalent antibodies (Fab-fragments of immunoglobulin G) is described in which Fab-fragments from sheep antiserum to rabbit Fab-fragments are first adsorbed to Sepharose containing insolubilized rabbit immunoglobulin G, and subsequently allowed to couple with ferritin previously activated with toluene-2,4-diisocyanate. The conjugate, after elution from the adsorbent, is separated from free ferritin and unconjugated Fab by gel chromatography. We have used this conjugate as a common second-step reagent for the surface localization of intracellular antigens on thin sections of tissue embedded in albumin. The first-step reagent consists of specific rabbit antibodies (IgG) directed against the tissue component under study. An example of the use of this technique for the localization of trypsinogen in bovine exocrine pancreatic cells is given.
关键词：immuno-electron microscopy ; protein embedment ; affinity chromatography ; trypsinogen ; bovine pancreas