A part of MilliporeSigma

17-459 | phospho-IRS1 (Ser307 mouse/ Ser312 Human) STAR ELISA Kit

96 assays  
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      Replacement Information

      Key Specifications Table

      Analytes AvailableSpecies ReactivityKey ApplicationsDetection Methods
      IRS1 H, M, R ELISA Chromogenic
      Catalogue Number17-459
      Brand Family Upstate
      Trade Name
      • STAR
      • Upstate
      Descriptionphospho-IRS1 (Ser307 mouse/ Ser312 Human) STAR ELISA Kit
      Alternate Names
      • Insulin Receptor Substrate
      Materials Required but Not Delivered1. Multi-channel or repeating pipettes
      2. Plate shaker (optional)
      3. Pipettors and 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
      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 IRS1 plate is coated with a specific mouse monoclonal capture antibody on the microwells of the 96-well clear plate. Sample lysate or the standard included in the kit are incubated in the microwells allowing IRS1 antigen 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-IRS1 (Ser312) antibody to detect the captured IRS1 on the plate well that is phosphorylated on Ser312. 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.

      IRS (Insulin Receptor Substrate) proteins are the effectors of both Insulin and IGF-initiated signaling. Insulin, in addition to working very similarly to a growth factor in stimulating receptor tyrosine kinases, is essential for maintaining glucose homeostasis and regulating carbohydrate, lipid, and protein metabolism. The signaling events of insulin are mediated via a vast and highly integrated network of molecules that controls several processes. There are 4 IRS proteins, 1-4. It is believed that these proteins serve complementary functions in different tissues. The two prominent players are IRS1 and IRS2 with IRS1 playing important roles in skeletal muscle and IRS2 in the liver. The IRS proteins link the insulin receptor with many signaling pathways including the PI3 Kinase/Akt/mTOR and the MAP Kinase pathways. The PI3K/Akt/mTOR pathway is involved with many metabolic actions of insulin signaling. MAPK is involved in the regulation of some genes and works with PI3 Kinase to control cell growth and differentiation. In addition to the insulin receptor tyrosine kinase activity in type II diabetes patients and mice, studies have shown a greater defect in components of the insulin signaling pathway including Insulin Receptor Substrate (IRS) phosphorylation or the activation of Phosphoinositide 3- Kinase (PI3 Kinase). Insulin stimulates the signaling cascade by binding to the insulin receptor through its extracellularα 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. IRS binds to the phosphorylated tyrosines of the β subunit of the insulin receptor. They share PH and PTB domains near their N-termini, and multiple Tyr phosphorylation motifs in their C-terminal regions. Proteins which bind to tyrosine-phosphorylated IRS-proteins include PI3 Kinase p85, GRB2, SHP2, Nck, Crk, and Fyn. IRS1 appears to be principally involved in IGF-signaling and cytoskeletal growth. IRS2 appears to be the critical mediator of Insulin signaling, as genetic ablation results in type II diabetes. IRS3 is expressed primarily in adipocytes and is an unusually potent activator of PI3 Kinase. IRS4 lacks the tyrosine residues which in the other IRS-proteins binds to SHP2.
      Product Information
      • Capture Plate pre-coated with anti-IRS1 antibody: (Part No. 17-459A) One pre-coated 96-stripwell immunoplate sealed in a foil pouch.
      • Anti-phospho-IRS1 (Ser312) detection antibody: (Part No. 17-459B) One bottle (11 mL) of anti- phospho- IRS1 (Ser312) detection antibody containing sodium azide, ready to use.
      • ELISA Diluent: (Part No. 17-459C) One bottle (25 mL) of ELISA Diluent containing sodium azide, ready to use.
      • 25X ELISA Wash Buffer: (Part No. 17-459D) One bottle (50 mL) of 25X ELISA Wash Buffer.
      • Anti-Rabbit IgG HRP conjugate: (Part No. 17-459E) One vial (125 μL) of 100X anti-rabbit HRP conjugate containing thimerosal.
      • HRP Diluent: (Part No. 17-459F) One bottle (25 mL) of HRP Diluent containing thimerosal.
      • TMB Solution: (Part No. 17-459G) One bottle (25 mL) of stabilized tetramethylbenzidine (TMB), ready to use.
      • Stop Solution: (Part No. 17-459H) One bottle (25 mL) of stop solution, ready to use.
      • Phospho-IRS1 (Ser312) Standard: (Part No. 17-459I) Two vials of IRS1 standard, lyophilized.
      • Plate Covers: Two plate covers.
      Detection methodChromogenic
      ApplicationThis phospho-IRS1 (Ser307 mouse/ Ser312 Human) STAR ELISA Kit is used to measure & quantify IRS1 levels.
      Key Applications
      • ELISA
      Biological Information
      Species Reactivity
      • Human
      • Mouse
      • Rat
      Analytes Available
      • IRS1
      Entrez Gene Number
      Gene Symbol
      • IRS1
      • IRS-1
      • HIRS-1
      • Phosphorylation
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: P35568 # May mediate the control of various cellular processes by insulin. When phosphorylated by the insulin receptor binds specifically to various cellular proteins containing SH2 domains such as phosphatidylinositol 3-kinase p85 subunit or GRB2. Activates phosphatidylinositol 3-kinase when bound to the regulatory p85 subunit (By similarity).
      SIZE: 1242 amino acids; 131591 Da
      SUBUNIT: Interacts with the NPXY motif of tyrosine-phosphorylated IGF1R and INSR via the PTB domain. Binds to phosphatidylinositol 3-kinase p85 subunit via the phosphorylated YXXM motifs. Binds ROCK1. Binds to UBTF and PIK3CA in nuclear extracts (By similarity). Interacts with SOCS7.
      PTM: Serine phosphorylation of IRS1 is a mechanism for insulin resistance. Ser-312 phosphorylation inhibits insulin action through disruption of IRS1 interaction with the insulin receptor (By similarity). & Phosphorylation of Tyr-896 is required for GRB2-binding (By similarity).DISEASE:SwissProt: P35568 # Polymorphisms in IRS1 may be involved in the etiology of non-insulin-dependent diabetes mellitus (NIDDM) [MIM:125853].
      SIMILARITY: SwissProt: P35568 ## Contains 1 IRS-type PTB domain. & Contains 1 PH domain.
      Physicochemical Information
      • Sensitivity: 1.5 Units/mL
        Range of Detection: 1.6 to 100 Units/mL
      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.


      • 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


      Certificates of Analysis

      TitleLot Number
      STAR phospho-IRS1 (Ser312) ELISA Kit - Manual Manual


      Reference overviewPub Med ID
      Tissue-specific insulin resistance in mice with mutations in the insulin receptor, IRS-1, and IRS-2
      Kido, Y, et al
      J Clin Invest, 105:199-205 (2000) 2000

      10642598 10642598
      Phosphoinositide 3-kinase: the key switch mechanism in insulin signalling
      Shepherd, P R, et al
      Biochem J, 333 ( Pt 3):471-90 (1998) 1998

      9677303 9677303
      Differential regulation of insulin receptor substrates-1 and -2 (IRS-1 and IRS-2) and phosphatidylinositol 3-kinase isoforms in liver and muscle of the obese diabetic (ob/ob) mouse
      Kerouz, N J, et al
      J Clin Invest, 100:3164-72 (1997) 1997

      9399964 9399964
      Insulin receptor phosphorylation, insulin receptor substrate-1 phosphorylation, and phosphatidylinositol 3-kinase activity are decreased in intact skeletal muscle strips from obese subjects
      Goodyear, L J, et al
      J Clin Invest, 95:2195-204 (1995) 1995

      7537758 7537758

      Technical Info

      96 well Template 2 Up
      96 well Template Single


      Multiplex Assays of Fifteen Human Complement Factors

      User Guides

      phospho-IRS1 (Ser312) STAR ELISA Kit
      phospho-IRS1 (Ser312) STAR ELISA Kit (Ser312 in human/Ser307 in mouse)

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