ADSORPTION OF HORSERADISH-PEROXIDASE, OVOMUCOID AND ANTIIMMUNOGLOBULIN TO COLLOIDAL GOLD FOR INDIRECT DETECTION OF CONCANAVALIN-A, WHEAT-GERM AGGLUTININ AND GOAT ANTIHUMAN IMMUNOGLOBULIN-G ON CELL-SURFACES AT ELECTRON-MICROSCOPIC LEVEL - NEW METHOD, THEORY AND APPLICATION
ADSORPTION OF HORSERADISH-PEROXIDASE, OVOMUCOID AND ANTIIMMUNOGLOBULIN TO COLLOIDAL GOLD FOR INDIRECT DETECTION OF CONCANAVALIN-A, WHEAT-GERM AGGLUTININ AND GOAT ANTIHUMAN IMMUNOGLOBULIN-G ON CELL-SURFACES AT ELECTRON-MICROSCOPIC LEVEL - NEW METHOD, THEORY AND APPLICATION
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DOI：
10.1177/25.11.21217
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发表时间：
1977-01-01
影响因子：
3.2
通讯作者：
ACKERMAN, GA
中科院分区：
文献类型：
--
作者：
GEOGHEGAN, WD;ACKERMAN, GA
描述了一种将选定的大分子吸附到胶体Au上的方法，然后将胶体Au用作间接检测特定细胞表面分子的电子致密标记物。用Au标记的辣根过氧化物酶和卵类粘蛋白分别间接检测膜结合伴刀豆球蛋白A和麦胚凝集素。用Au标记的兔抗山羊IgG检测血淋巴细胞上的山羊抗人IG[免疫球蛋白]M。在胶体Au标记蛋白的制备中，蛋白质对胶体Au的絮凝和蛋白质不吸附到胶体金的问题通过蛋白质浓度和pH可变吸附等温线的组合来解决，这允许人们确定蛋白质吸附到胶体Au的条件。吸附是pH依赖性的，pH条件与主要蛋白质组分的等电点相关;吸附受界面张力、溶解度和分子上的电荷的影响。胶体Au是廉价的，并且制备有用的标记物是快速的、可重复的，并且结果容易从电子显微照片定量。
A method is described for the adsorption of selected macromolecules to colloidal Au which is then used as an electron dense marker for the indirect detection of specific cell surface molecules. Membrane bound concanavalin A, which binds specific sugars on horseradish peroxidase, and wheat germ agglutinin, which binds specific sugars on ovomucoid are detected indirectly with Au labeled horseradish peroxidase and ovomucoid, respectively. Goat anti-human Ig[immunoglobulin]M on blood lymphocytes is detected with Au labeled rabbit anti-goat IgG. In the preparation of colloidal Au labeled proteins, the problems of flocculation of colloidal Au by proteins and nonadsorption of proteins to colloidal gold, are solved through a combination of concentration of protein and pH variable adsorption isotherms, which allows one to determine the conditions for adsorption of proteins to colloidal Au. Adsorption is pH dependent, the pH conditions correlating with the isoelectric point(s) of the major protein fraction(s); adsorption is influenced by interfacial tension, solubility and by the electrical charge on the molecules. Colloidal Au is inexpensive and preparation of a useful label is rapid, reproducible and the results easily quantitated from electron micrographs.