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phospho-Insulin Receptor, β subunit STAR ELISA Assay Kit
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PRODUCT FAMILY INFORMATION
STAR ELISA Kits
Upstate STAR (Signal Transduction Assay Reaction) ELISA kits are a fast, sensitive method to detect the relative amounts of total and activated targets through the use of phospho-specific antibodies.
Upstate STAR (Signal Transduction Assay Reaction) ELISA kits are a fast, sensitive method to detect the relative amounts of total and activated targets through the use of phospho-specific antibodies.
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Description:
phospho-IR (β Subunit) (Tyr 1162/1163) STAR ELISA Assay Kit
Trade Name:
STAR ELISA
Upstate (Millipore)
Qty/Pk:
96 assays
Background Information:
I. TEST PRINCIPLE
The UPSTATE® colorimetric STAR (Signal Transduction Assay Reaction) ELISA kit is a solid phase sandwich enzyme linked immunosorbent assay that provides a fast, sensitive method to detect specific levels of signaling targets in whole cell extracts. The IR plate is coated with a specific mouse monoclonal IR (β subunit) capture antibody on the microwells of the 96-well clear plate. Sample lysate or the standards included in the kit are incubated in the microwells allowing IR to be captured in the plate wells. The plate is then washed to remove any unbound non-specific material. The wells are then incubated with a specific rabbit anti-phospho-IR antibody to detect the captured IR phosphorylated on Tyrosines 1162 and 1163. The unbound detection antibody is washed away followed by incubation with an HRP-conjugated anti-rabbit antibody. This allows for a sensitive enzymatic detection of the sample. After the addition of TMB substrate and stop solution the absorbance is measured at 450 nm using a plate reader.
The entire assay takes less than 5 hours to complete with minimal hands-on time. Many of the reagents are supplied in ready-to use formulations for ease of use. The kit also includes a standard that is run as both a positive control and to develop a standard curve.
II. IR BACKGROUND
The Insulin Receptor (IR) is synthesized as a single polypeptide, which is subsequently cleaved to generate an extracellular α chain and a transmembrane and intracellular β chain, tethered together by disulfide bonds. The β chain has multiple tyrosine phosphorylation sites, including three autophosphorylation sites at its activation loop. Insulin stimulates the signaling cascade by binding to the insulin receptor through its extracellular α subunits. This stimulates the tyrosine kinase activity of the β subunits of the receptor. The receptor then autophosphorylates itself and phosphorylates the IRS (Insulin Receptor Substrate) proteins that are very important modulators of insulin signaling. Phosphorylated IRS, in turn, interacts with several downstream effectors of insulin signaling, including PI3 Kinase p85, GRB2, SHP2, Nck, Crk, and Fyn. The overall structure of the IR is highly homologous to the IGF-I Receptor, except in their c-termini, where the two proteins diverge somewhat. Insulin signaling is highly dependent on the PI3 Kinase pathway and Akt, which appear to mediate the functions of insulin.
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Applications:
This phospho-IR (β Subunit) (Tyr 1162/1163) STAR ELISA Assay Kit is used to measure & quantify Insulin levels.
Key Applications:
ELISA
Species Reactivity:
Human
Mouse
Rat
Usage Statement:
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animal |