The Human IR (Phospho) [pY1158] ELISA Kit is a solid-phase sandwich Enzyme-Linked Immunosorbent Assay (ELISA) designed to detect and quantify the level of IR (Phospho) [pY1158] in fresh or frozen human cell lysates. Cross-reactivity with mouse and rat cells has been observed. The assay recognizes both natural and recombinant human IR (Phospho) [pY1158].
Principle of the method
A monoclonal capture antibody specific for IR (Phospho) [pY1158] has been coated onto the wells of the 96-well plate. During the first incubation, standards of known content and unknown samples are pipetted into the wells and the antigen binds to the immobilized (capture) antibody. After washing, a rabbit antibody specific for the target protein is added to the wells and serves as a detection antibody by binding to the immobilized protein captured during the first incubation. After washing, a horseradish peroxidase labeled anti-rabbit IgG is added. This binds to the detection antibody to complete the four member sandwich. After a third incubation and washing to remove all the 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 target protein present in the original specimen and the optical density can be read on a standard microplate reader.
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.
INSR (insulin receptor, IR) is a heterodimeric protein complex that has an intracellular beta subunit and an extracellular alpha subunit, which is disulfide- linked to a transmembrane segment. The insulin ligand binds to the IR and initiates molecular signaling pathways that promote glucose uptake in cells and glycogen synthesis. Insulin binding to IR induces phosphorylation of intracellular tyrosine kinase domains and recruitment of multiple SH2 and SH3 domain-containing intracellular proteins that serve as signaling intermediates for pleiotropic effects of insulin. INSR and IGF-1 receptors share major structural and functional similarity. The earliest cellular response to insulin stimulation is autophosphorylation of tyrosine in INSR. In humans, the INSR gene is located on chromosome 19. Defects in INSR are the cause of various insulin resistance syndromes and IGF-1R defects may also cause some forms of growth retardation.
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