Revision 6
#7258
Store at +4C
PathScan® Phospho-Insulin Receptor β (Tyr1150/1151) Sandwich ELISA Kit
1 Kit
(96 assays)
Species Cross Reactivity:
H M
UniProt ID:
#P06213
Entrez-Gene Id:
#3643
877-616-CELL (2355)
877-678-TECH (8324)
3 Trask Lane | Danvers | Massachusetts | 01923 | USA
For Research Use Only. Not for Use in Diagnostic Procedures.
| Product Includes | Product # | Quantity | Color | Storage Temp |
|---|---|---|---|---|
| Insulin Receptor β Mouse mAb Coated Microwells | 70302 | 96 tests | +4C | |
| Phospho-Insulin Receptor β (Tyr1150/1151) Rabbit Detection mAb | 12911 | 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
CST's PathScan® Phospho-Insulin Receptor β (Tyr1150/1151) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects transfected phospho-insulin receptor (Tyr1150/1151) protein. An Insulin Receptor β Mouse mAb has been coated onto the microwells. After incubation with cell lysates, both phospho- and nonphospho-insulin receptor proteins are captured by the coated antibody. Following extensive washing, Phospho-IGF-I Receptor β (Tyr1135/1136)/Insulin Receptor β (Tyr1150/1151) Rabbit mAb is added to detect the captured phospho-insulin receptor (Tyr1150/1151) 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 optical density for this developed color is proportional to the quantity of phospho-insulin receptor β (Tyr1150/1151) protein.
*Antibodies in kit are custom formulations specific to kit.
Specificity/Sensitivity
CST's PathScan® Phospho-Insulin Receptor β (Tyr1150/1151) Sandwich ELISA Kit #7258 detects phospho-insulin receptor (Tyr1150/1151) protein. As shown in Figure 1, using Phospho-Insulin Receptor β (Tyr1150/1151) Sandwich ELISA Kit #7258, a significant induction of phospho-insulin receptor (Tyr1150/1151) is detected in CHO-IR/IRS-1 cells treated with insulin. The levels of total insulin receptor β (phospho and nonphospho) shown by Western analysis remain unchanged. Endogenous phospho-insulin receptor β (Tyr1150/1151) in either NIH/3T3 or HepG2 cells treated with insulin is also detected by Phospho-Insulin Receptor β (Tyr1150/1151) Sandwich ELISA Kit #7258. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
Background
Type I insulin-like growth factor receptor (IGF-IR) is a transmembrane receptor tyrosine kinase that is widely expressed in many cell lines and cell types within fetal and postnatal tissues (1-3). Receptor autophosphorylation follows binding of the IGF-I and IGF-II ligands. Three tyrosine residues within the kinase domain (Tyr1131, Tyr1135, and Tyr1136) are the earliest major autophosphorylation sites (4). Phosphorylation of these three tyrosine residues is necessary for kinase activation (5,6). Insulin receptors (IRs) share significant structural and functional similarity with IGF-I receptors, including the presence of an equivalent tyrosine cluster (Tyr1146/1150/1151) within the kinase domain activation loop. Tyrosine autophosphorylation of IRs is one of the earliest cellular responses to insulin stimulation (7). Autophosphorylation begins with phosphorylation at Tyr1146 and either Tyr1150 or Tyr1151, while full kinase activation requires triple tyrosine phosphorylation (8).
Background References
- Adams, T.E. et al. (2000) Cell Mol Life Sci 57, 1050-93.
- Baserga, R. (2000) Oncogene 19, 5574-81.
- Scheidegger, K.J. et al. (2000) J Biol Chem 275, 38921-8.
- Hernández-Sánchez, C. et al. (1995) J Biol Chem 270, 29176-81.
- Lopaczynski, W. et al. (2000) Biochem Biophys Res Commun 279, 955-60.
- Baserga, R. (1999) Exp Cell Res 253, 1-6.
- White, M.F. et al. (1985) J Biol Chem 260, 9470-8.
- White, M.F. et al. (1988) J Biol Chem 263, 2969-80.
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 G: Goat 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 6
Figure 1: Treatment of CHO-IR/IRS-1 cells with insulin stimulates phosphorylation of insulin receptor at Tyr1150/1151, detected by PathScan® Phospho-Insulin Receptor β (Tyr1150/1151) Sandwich ELISA Kit #7258, but the level of total insulin receptor shown by Western analysis remains unchanged. OD 450 nm eadings are shown in the top figure, while the 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 Antibody #3025 (left panel), are shown in the bottom figure. CHO-IR/IRS-1 cells stably overexpress human insulin receptor and rat IRS-1.
Figure 2: The relationship between protein concentration of lysates from untreated and insulin-treated CHO-IR/IRS-1 cells and kit assay optical density readings 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.
