Revision 5

#7069Store at +4C

1 Kit

(96 assays)

Species Cross Reactivity

H

UniProt ID:

#P06213

Entrez-Gene Id:

#3643

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Color Storage Temp
Insulin Receptor β Mouse mAb Coated Microwells 86530 96 tests +4C
Insulin Receptor beta Rabbit Detection mAb 5858 1 ea Green (Lyophilized) +4C
Anti-rabbit IgG, HRP-linked Antibody (ELISA Formulated) 13272 1 ea Red (Lyophilized) +4C
Detection Antibody Diluent 13339 11 ml Green +4C
HRP Diluent 13515 11 ml Red +4C
TMB Substrate 7004 11 ml +4C
STOP Solution 7002 11 ml +4C
Sealing Tape 54503 2 ea +4C
ELISA Wash Buffer (20X) 9801 25 ml +4C
ELISA Sample Diluent 11083 25 ml Blue +4C
Cell Lysis Buffer (10X) 9803 15 ml -20C

*The microwell plate is supplied as 12 8-well modules - Each module is designed to break apart for 8 tests.

Description

PathScan® Total Insulin Receptor β Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects transfected levels of insulin receptor β protein. An Insulin Receptor β Mouse mAb has been coated on the microwells. After incubation with cell lysates, insulin receptor β protein (phospho and nonphospho) is captured by the coated antibody. Following extensive washing, an Insulin Receptor β Rabbit mAb is added to detect captured Insulin Receptor β protein. Anti-rabbit IgG, HRP-linked antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of the absorbance for this developed color is proportional to the quantity of insulin receptor β protein.

*Antibodies in this kit are custom formulations specific to kit.

Specificity/Sensitivity

PathScan® Total Insulin Receptor β Sandwich ELISA Kit #7069 detects transfected levels of human Insulin Receptor β protein, as shown in Figure 1. The kit sensitivity is shown in Figure 2. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Background

Insulin receptor (INSR) is a membrane receptor tyrosine kinase. The receptor molecule consists of a disulfide linked heterodimer. The α subunit is a 135 kDa extracellular fragment, and the β subunit is a 95 kDa fragment containing an extracellular domain, a single transmembrane domain, and an intracellular tyrosine kinase domain (1). Insulin ligand binding to this receptor results in receptor autophosphorylation and tyrosine kinase activation. INSR catalyzes the tyrosine phosphorylation of molecules such as IRS, Gab1, Shc, and Cbl, which further activate the downstream MAPK, PI3K, and TC10 pathways. This eventually leads to increases in glucose uptake and metabolism as well as cell growth (2,3). INSR has peptide substrate specificity similar to other receptor tyrosine kinase members, preferring acidic residues at the -1 to -4 positions and large hydrophobic amino acids at positions +1 and +3 (4).

  1. Yip, C.C. and Ottensmeyer, P. (2003) J Biol Chem 278, 27329-32.
  2. Saltiel, A.R. and Pessin, J.E. (2002) Trends Cell Biol 12, 65-71.
  3. Zick, Y. (2001) Trends Cell Biol 11, 437-41.
  4. Songyang, Z. and Cantley, L.C. (1995) Trends Biochem Sci 20, 470-5.

Background References

    Cross-Reactivity Key

    H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    PathScan is a registered trademark of Cell Signaling Technology, Inc.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

    Limited Uses

    Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

    Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

    Revision 5
    #7069

    PathScan® Total Insulin Receptor β Sandwich ELISA Kit

    PathScan® Total Insulin Receptor β Sandwich ELISA Kit: Image 1 Expand Image
    Figure 1: Treatment of CHO-IR/IRS-1 cells with insulin stimulates tyrosine phosphorylation of Insulin Receptor β which is detected by PathScan® Phospho-Insulin Receptor β (panTyr) Sandwich ELISA Kit #7082 (top, right). Similar levels of Insulin Receptor β protein from both nonphospho or phospho lysates are detected by PathScan® Total Insulin Receptor β Sandwich ELISA Kit #7069 (top, left). Absorbance at 450 nm is shown in the top figure while corresponding western blots using Phospho-IGF-I Receptor β (Tyr1135/1136)/Insulin Receptor β (Tyr1150/1151) (19H7) Rabbit mAb #3024 (right panel) or Insulin Receptor β (4B8) Rabbit mAb #3025 (left panel) are shown in the bottom figure. CHO-IR/IRS-1 cells stably overexpress the tranfected human insulin receptor and rat IRS-1.
    PathScan® Total Insulin Receptor β Sandwich ELISA Kit: Image 2 Expand Image
    Figure 2: The relationship between protein concentration of lysates from untreated and insulin-treated CHO-IR/IRS-1 cells and the absorbance at 450 nm is shown. After starvation, CHO-IR/IRS-1 cells (85% confluence) were treated with insulin (100 nM) for 2 min at 37°C and then lysed.