This IR (beta-subunit) ELISA is designed to detect and quantify levels of the insulin receptor b-subunit independent of its phosphorylation status. Both natural (heterotetrameric) and recombinant IR react in this assay. This ELISA is not cross-reactive with IGF-1R. Although this ELISA kit is developed using cells containing human insulin receptor, mouse and rat insulin receptor will cross react. This assay is intended for detection of IR from lysates of cells.
This kit is a solid phase sandwich Enzyme Linked-Immuno-Sorbent Assay (ELISA). A monoclonal antibody specific for IR (b-subunit) (regardless of phosphorylation state) has been coated onto the wells of the microtiter strips provided. Samples, including a standard containing IR, control specimens, and unknowns, are pipetted into these wells. During the first incubation, the IR (b-subunit) antigen binds to the immobilized (capture) antibody. After washing, an antibody specific for IR (b-subunit) is added to the wells. During the second incubation, this antibody serves as a detection antibody by binding to the immobilized IR (b-subunit) protein captured during the first incubation. After removal of excess detection antibody, a horseradish peroxidase-labeled Anti-Rabbit IgG (Anti-Rabbit IgG HRP) is added. This binds to the detection antibody to complete the four-member sandwich. After a third incubation and washing to remove all the excess Anti-Rabbit IgG HRP, a substrate solution 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 IR (b-subunit) present in the original specimen.
Sensitivity:0.5ng/ml
Range:0.94-60ng/ml
Incubation Time:4 hours
Sample size:100ul of sample diluted by a factor 1:10 or greater.
Economical:One 96-well plate (12x9 wells trips) plus reagents for 96 determinations
Storage and Stability:Store all components at 4°C. Stable for 6 months. For maximum recovery of product, centrifuge the original vial prior to removing the cap.
