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17-483 Insulin Receptor (β subunit) STAR ELISA Assay Kit

This Insulin Receptor (β subunit) STAR ELISA Assay Kit is used to measure & quantify Insulin levels.

MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
Key Applications: ELISA Detection Methods: Chromogenic Research Category: Epigenetics & Nuclear Function, Cell Signaling
17-483
96 assays  
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      Overview

      Replacement Information

      Key Spec Table

      Analytes AvailableSpecies ReactivityKey ApplicationsDetection Methods
      Insulin H, M, RELISAChromogenic
      Description
      Catalogue Number17-483
      Brand Family Chemicon®
      Trade Name
      • STAR
      • Chemicon
      DescriptionInsulin Receptor (β subunit) STAR ELISA Assay Kit
      Alternate Names
      • Insulin Receptor
      • IR
      Background InformationI. 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-IR antibody to detect the captured IR (β subunit) on the plate well. 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.
      Materials Required but Not Delivered1. Multi-channel or repeating pipettes 2. Plate shaker (optional) 3. Pipettors & tips capable of accurately measuring 1-1000 µL 4. Graduated serological pipettes 5. 96-well microtiter Plate Reader with 450 nm filter 6. Graphing software for plotting data or graph paper for manual plotting of data 7. Microfuge tubes for standard and sample dilutions 8. Mechanical vortex 9. 1 liter container 10. Distilled or deionized water
      References
      Product Information
      Components
      • Capture Plate pre-coated with anti-IR antibody: (Part No. 17-483A) One pre-coated 96-stripwell immunoplate sealed in a foil pouch.
      • Anti-IR detection antibody: (Part No. 17-483B) One bottle (11 mL) of anti-IR (β subunit) detection antibody containing sodium azide, ready to use. .
      • ELISA Diluent: (Part No. 17-483C) One bottle (25 mL) of ELISA Diluent containing sodium azide, ready to use.
      • 25X ELISA Wash Buffer: (Part No. 17-483D) One bottle (50 mL) of 25X ELISA Wash Buffer.
      • Anti-Rabbit IgG HRP conjugate: (Part No. 17-483E) One vial (125 µL) of 100X anti-rabbit HRP conjugate containing thimerosal.
      • HRP Diluent: (Part No. 17-483F) One bottle (25 mL) of HRP Diluent containing thimerosal.
      • TMB Solution: (Part No. 17-483G) One bottle (25 mL) of stabilized tetramethylbenzidine (TMB), ready to use.
      • Stop Solution: (Part No. 17-483H) One bottle (25 mL) of stop solution, ready to use.
      • IR Standard: (Part No. 17-483I) Four vials of IR (β subunit) standard, lyophilized
      • Plate Covers: Two plate covers.
      Detection methodChromogenic
      Applications
      ApplicationThis Insulin Receptor (β subunit) STAR ELISA Assay Kit is used to measure & quantify Insulin levels.
      Key Applications
      • ELISA
      Biological Information
      Species Reactivity
      • Human
      • Mouse
      • Rat
      Analytes Available
      • Insulin
      Entrez Gene Number
      Entrez Gene SummaryAfter removal of the precursor signal peptide, the insulin receptor precursor is post-translationally cleaved into two chains (alpha and beta) that are covalently linked. Binding of insulin to the insulin receptor (INSR) stimulates glucose uptake. Two transcript variants encoding different isoforms have been found for this gene.
      Gene Symbol
      • INSR
      • CD220
      • IR
      • HHF5
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: P06213 # This receptor binds insulin and has a tyrosine-protein kinase activity. Isoform Short has a higher affinity for insulin. Mediates the metabolic functions of insulin. Binding to insulin stimulates association of the receptor with downstream mediators including IRS1 and phosphatidylinositol 3'-kinase (PI3K). Can activate PI3K either directly by binding to the p85 regulatory subunit, or indirectly via IRS1.
      SIZE: 1382 amino acids; 156307 Da
      SUBUNIT: Tetramer of 2 alpha and 2 beta chains linked by disulfide bonds. The alpha chains contribute to the formation of the ligand- binding domain, while the beta chains carry the kinase domain. Interacts with SORBS1 but dissociates from it following insulin stimulation. Binds SH2B2. Interacts with the PTB/PID domains of IRS1 and SHC1 in vitro when autophosphorylated on tyrosine residues. The sequences surrounding the phosphorylated NPXY motif contribute differentially to either IRS1 or SHC1 recognition. Interacts with the SH2 domains of the 85 kDa regulatory subunit of PI3K (PIK3R1) in vitro, when autophosphorylated on tyrosine residues. Interacts with SOCS7.
      SUBCELLULAR LOCATION: Membrane; Single-pass type I membrane protein.
      TISSUE SPECIFICITY: Isoform Long and isoform Short are expressed in the peripheral nerve, kidney, liver, striated muscle, fibroblasts and skin. Isoform Short is expressed also in the spleen and lymphoblasts.
      PTM: After being transported from the endoplasmic reticulum to the Golgi apparatus, the single glycosylated precursor is further glycosylated and then cleaved, followed by its transport to the plasma membrane. & Autophosphorylated on tyrosine residues in response to insulin. & Phosphorylation of Tyr-999 is required for IRS1- and SHC1- binding.
      DISEASE: SwissProt: P06213 # Defects in INSR are the cause of insulin resistance (Ins resistance) [MIM:125853]. & Defects in INSR are the cause of Rabson-Mendenhall syndrome [MIM:262190]; also known as Mendenhall syndrome. It is a severe insulin resistance syndrome characterized by insulin- resistant diabetes mellitus with pineal hyperplasia and somatic abnormalities. Typical features include coarse, senile-appearing facies, dental and skin abnormalities, abdominal distension, and phallic enlargement. Inheritance is autosomal recessive. & Defects in INSR are the cause of leprechaunism [MIM:246200]; also known as Donohue syndrome. Leprechaunism represents the most severe form of insulin resistance syndrome, characterized by intrauterine and postnatal growth retardation and death in early infancy. Inheritance is autosomal recessive. & Defects in INSR may be associated with noninsulin- dependent diabetes mellitus (NIDDM) [MIM:125853]; also known as diabetes mellitus type 2. & Defects in INSR are the cause of familial hyperinsulinemic hypoglycemia 5 (HHF5) [MIM:609968]. Familial hyperinsulinemic hypoglycemia [MIM:256450], also referred to as congenital hyperinsulinism, nesidioblastosis, or persistent hyperinsulinemic hypoglycemia of infancy (PPHI), is the most common cause of persistent hypoglycemia in infancy and is due to defective negative feedback regulation of insulin secretion by low glucose levels. & Defects in INSR are the cause of insulin-resistant diabetes mellitus with acanthosis nigricans type A (IRAN type A) [MIM:610549]. This syndrome is characterized by the association of severe insulin resistance (manifested by marked hyperinsulinemia and a failure to respond to exogenous insulin) with the skin lesion acanthosis nigricans and ovarian hyperandrogenism in adolescent female subjects. Women frequently present with hirsutism, acne, amenorrhea or oligomenorrhea, and virilization. This syndrome is different from the type B that has been demonstrated to be secondary to the presence of circulating autoantibodies against the insulin receptor.
      SIMILARITY: SwissProt: P06213 ## Belongs to the protein kinase superfamily. Tyr protein kinase family. Insulin receptor subfamily. & Contains 2 fibronectin type-III domains. & Contains 1 protein kinase domain.
      Physicochemical Information
      Sensitivity
      • Sensitivity: 0.75 ng/mL
        Range of Detection: 0.94 to 60 ng/mL
        Species Reactivity: Human, mouse and rat
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality AssuranceRoutinely evaluated by ELISA
      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 animals.
      Storage and Shipping Information
      Storage ConditionsMaintain the unopened kit at 2-8°C until expiration date.

      Precautions

      • The instructions provided have been designed to optimize the kit's performance. Deviation from the instructions may result in suboptimal performance of the kit and the failure to produce accurate data.
      • Caustic Material: Stop Solution. Caution: Eye, hand, face, and clothing protection should be worn when handling this material.
      • Safety Warnings and Precautions: This kit is designed for research use only and not recommended for internal use in humans or animals. All chemicals should be considered potentially hazardous and principles of good laboratory practice should be followed.
      • The Detection Antibody and ELISA Diluent contain sodium azide. Sodium azide may react with copper and lead plumbing to form highly explosive metal azides. Upon disposal, flush with large amounts of water to prevent azide build-up. Avoid contact with skin.
      • The Anti-Rabbit IgG HRP Conjugate and HRP Diluent contain thimerosal. Thimerosal is highly toxic by inhalation, contact with skin or if swallowed. Thimerosal is a possible mutagen and should be handled accordingly.
      Packaging Information
      Material Size96 assays
      Transport Information
      Supplemental Information
      Specifications
      Global Trade ITEM Number
      Catalogue Number GTIN
      17-483 04053252330056

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