Invitrogen™ Rat Ig Isotyping Instant ELISA™ Kit 

Description

The Rat Immunoglobulin Isotyping ELISA qualitatively detects Rat Ig isotypes in rat hybridoma cell culture supernatants.

Principle of the method Biotinylated monoclonal anti-rat isotype specific or light chain specific capture antibodies bind to the Streptavidin coated plate. Ig Isotype present in the positive control or sample binds to the specific capture antibodies and is simultaneously bound by the HRP-conjugated detector antibody. Following incubation unbound reagents and sample components are removed during a wash step, and substrate solution reactive with HRP is added to the wells. A colored product is formed in proportion to the amount of soluble rat Ig (H+L) present in the sample.

Rigorous validation Each manufactured lot of this ELISA kit is quality tested. See manual for more information on validation.

The isotype of a primary antibody and the application it is being used in can result in background staining. Primary antibody background noise can be caused by binding to Fc receptors on target cells; by non-specific interactions with cellular proteins, carbohydrates, and lipids; or by cell autofluorescence. Isotype control antibodies can act as negative controls to help differentiate non-specific background signal from specific antibody signal because they have no relevant specificity to a target antigen. While isotype controls are most commonly used in flow cytometry, they are useful in other applications such as chromatin immunoprecipitation (ChIP), immunohistochemistry, and gel shifts. Isotype controls should match with the primary antibody species and isotype so that the level of specific staining by the primary antibody may be accurately determined. If using directly labeled primary antibodies, the isotype control works best if conjugated with the same label as the test antibody.