Invitrogen™ IFN gamma Human ELISA Kit

Description

The Human Interferon-gamma (Hu IFN-γ) ELISA quantitates Hu IFN-γ in human serum, buffered solution, or cell culture medium. The assay will exclusively recognize both natural and recombinant Hu IFN-γ. Principle of the method The Human IFN-γ solid-phase sandwich ELISA (enzyme-linked immunosorbent assay) is designed to measure the amount of the target bound between a matched antibody pair. A target-specific antibody has been pre-coated in the wells of the supplied microplate. Samples, standards, or controls are then added into these wells and bind to the immobilized (capture) antibody. The sandwich is formed by the addition of the second (detector) antibody, a substrate solution is added that reacts with the enzyme-antibody-target complex to produce measurable signal. The intensity of this signal is directly proportional to the concentration of target present in the original specimen. Rigorous validation Each manufactured lot of this ELISA kit is quality tested for criteria such as sensitivity, specificity, precision, and lot-to-lot consistency. See manual for more information on validation.

Detects and quantifies the level of human IFN-γ in serum and supernatant using 96-well plates and a microplate reader. Convenient precoated, 96-well plate supplied as removable 8-well strips to accommodate smaller samples run. Easy-to-run sandwich ELISA has a total assay incubation time is only 2.75 hours. A monoclonal capture antibody specific for Human IFN-γ machine coated onto the wells of the 96-well plate, ensuring low well-to-well variability and eliminating the need for overnight coating. Samples, including a standard of known Human IFN-γ content, controls, and unknowns, are pipetted into these wells followed by an addition of a biotinylated polyclonal detector antibody. During the first incubation, Human IFN-γ antigen binds simultaneously to the immobilized capture antibody and to the solution phase biotinylated detector antibody on a second site. After washing, a streptavidin-horseradish peroxidase (HRP) is added. This binds to the biotinylated detection antibody to complete the four–member sandwich. After a second incubation and washing to remove all unbound enzyme, a substrate solution (TMB) is added, which is acted upon by the bound enzyme to produce color. The intensity of this colored product is directly proportional to the concentration of Human IFN-γ present in the original specimen and the optical density can be read on a standard microplate reader.

See protocol insert for more information on validation.

Cell Analysis, Cell Signaling, Cell Viability, Proliferation & Function, Immunoassays, Ready-To-Use Immunoassay

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