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Sensitive alkaline phosphatase linked secondary antibody system for detection of group specific component after isoelectric focusing on 250 μm thick reusable immobilized pH‐gradients
Author(s) -
Pflug Werner
Publication year - 1986
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150070606
Subject(s) - nitrocellulose , chromatography , isoelectric focusing , chemistry , immunofixation , cellulose acetate , immobilized ph gradient , cellulose , alkaline phosphatase , membrane , antibody , biochemistry , monoclonal antibody , enzyme , biology , monoclonal , immunology
A sensitive secondary antibody system was developed for detection of small amounts of group specific component (Gc). For the first antibody step after focusig, a comparison between immunofixation on cellulose acetate membranes and capillary blotting on nitrocellulose was made. The sensitivity of the first is much better than that of the latter. The method allowed the detection of about 150 picograms of Gc in 1:12 800 diluted serum samples. Even in six‐year‐old badly soluble bloodstains kept at room temperature, Gc could be phenotyped. The 250 μm thick immobilized pH‐gradient gels for focusing of Gc can be used repeatedly after washing and drying to the initial weight without destroying the pH gradient.