Description
This assay employs a two-site sandwich ELISA to quantitate IFN-alpha in samples. An antibody specific for IFN-alpha has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any IFN-alpha present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for IFN-alpha is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of IFN-alpha bound in the initial step. The color development is stopped and the intensity of the color is measured.
Overview: Streuli et al. (1980) showed that at least 3 different IFN-alpha genes are expressed in man. Furthermore, study of genomic DNA revealed the presence of at least 8 IFN genes. Nagata et al. (1980) found that the alpha-interferon genes are devoid of intervening sequences. Using radioactive probes from purified cDNA clones of interferons, Owerbach et al. (1981) located at least 8 leukocyte interferon genes and a fibroblast interferon gene on chromosome 9. Shows et al. (1982) found that the alpha- and beta-interferon genes are on 9p. The mapping to 9pter-q12 was accomplished by blot hybridization of cloned interferon cDNA to DNA from human-mouse cell hybrids with a translocation involving chromosome 9