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Description

The Hybridoma Subisotyping Kit is intended for the detection and identification of mouse monoclonal antibody subclasses IgG1, IgG2a, IgG2b, IgM and IgA in supernatants. It is also used for confirmation of monoclonality of cloned hybridoma cultures. This double antibody detection system is a highly sensitive enzyme immunoassay method incorporating specific primary antisera. Assay sensitivity is less than 1 µ,g/ml and can be reliably used with diluted culture supernatants. The following protocol utilizes 50 µ,l X 7 (350 µ,l) of each supernatant tested (a 1:2 dilution occurs as each sample is added to diluent in each well). When supernatant medium is precious, the medium from high producer cells can be diluted much more (1:10 or greater), with appropriate modifications in the Application of Supernatants section. When used as directed, the reagents in this kit will be sufficient for 900 determinations or 150 clones, Useful to confirm the purity of cell lines in monoclonal antibody development assays. Sufficient to perform 900 assays (150 clones).