Invitrogen™ TNF alpha Human ELISA Kit

Description

The Human Tumor Necrosis Factor alpha (Hu TNFα) ELISA quantitates Hu TNFα in human serum, plasma, buffered solution, or cell culture medium. The assay will exclusively recognize both natural and recombinant Hu TNFα. Principle of the method The Human TNFα 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.

Research use only kit designed to detect and quantify the level of human TNF-α in serum, plasma, and supernatant using 96-well plates and a microplate reader.

Easy-to-Run Sandwich ELISA
A monoclonal capture antibody specific for Human TNF-α has been 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 TNF-α content, controls, and unknowns, are pipetted into these wells. During the first incubation, Human TNF-α antigen binds to the immobilized capture antibody on one site. After washing, a biotinylated monoclonal antibody specific for Human TNF-α is added. During the second incubation, this antibody binds to the immobilized Human TNF-α antigen at a different 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 TNF-α present in the original specimen and the optical density can be read on a standard microplate reader. Total assay incubation time is only four hours.

• Easy to run
• Rapid protocol
• Convenient precoated, 96-well plate supplied as removable 8-well strips to accommodate smaller samples run

See protocol insert for more information on validation.

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

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