MICROFLUOROMETRIC IMMUNOASSAYS WITH ANTIGENS BOUND TO SEPHAROSE BEADS
| Autor: | C. M. Ham, F. J. Bloemmen, J. J. Haaijman |
|---|---|
| Rok vydání: | 1975 |
| Předmět: |
Ovalbumin
Fluorescent Antibody Technique Binding Competitive Antibodies General Biochemistry Genetics and Molecular Biology Sepharose Mice History and Philosophy of Science Antigen Polysaccharides Animals Humans Antigens Binding site Radial immunodiffusion Antiserum Chromatography biology Chemistry Immune Sera General Neuroscience Radioimmunoassay Fluorescence Immunoglobulin M Ethanolamines Immunoglobulin G biology.protein Rabbits |
| Zdroj: | Annals of the New York Academy of Sciences. 254:137-149 |
| ISSN: | 1749-6632 0077-8923 |
| Popis: | Sepharose beads, to which protein had been coupled, were used in the development of two types of microfluorometric immunoassays. The first is an inhibition assay: a fixed amount of antiserum was absorbed with different amounts of antigen. This was accomplished in both direct and indirect incubation schemes. The system has been studied with human IgG and mouse IgM as an example. The second is a competition assay: fluorescein-labeled ovalbumin competea with unlabeled unlabeled ovalbumin for the binding sites of a rabbit antiovalbumin. This antiovalbumin was not bound directly to Sepharose but to Sepharose-coupled ovalbumin. In the indirect fluorescence inhibition and the competition assay, the sensitivity levels were determined. They proved to be the same, namely, 25 ng. This means that these fluorescence immunoassays are 10 times less sensitive than conventional radioimmunoassays and 10 times more sensitive than conventional single radial immunodiffusion. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Plný text