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PathScan® Sandwich ELISA Kit

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Product Includes Volume Solution Color
Insulin Receptor beta Mouse mAb coated microwells 96 tests
Phospho-Insulin Receptor β (Tyr1150/1151) Rabbit Detection mAb 1 Ea Green (Lyophilized)
Anti-rabbit IgG, HRP-linked Antibody (ELISA Formulated) 1 Ea Red (Lyophilized)
Detection Antibody Diluent 11 ml Green
HRP Diluent 11 ml Red
TMB Substrate 7004 11 ml Colorless
STOP Solution 7002 11 ml Colorless
ELISA Wash Buffer (20X) 25 ml Colorless
ELISA Sample Diluent 25 ml Blue
Sealing Tape 2 sheets
Cell Lysis Buffer (10X) 9803 15 ml Yellowish

Product 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.

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).

1.  Adams, T.E. et al. (2000) Cell. Mol. Life Sci. 57, 1050-1093.

2.  Baserga, R. et al. (2000) Oncogene 19, 5574-5581.

3.  Scheidegger, K.J. et al. (2000) J. Biol. Chem. 275, 38921-38928.

4.  Hernandez-Sanchez, C. et al. (1995) J. Biol. Chem. 270, 29176-29181.

5.  Lopaczynski, W. et al. (2000) Biochem. Biophys. Res. Commun. 279, 955-960.

6.  Baserga, R. et al. (1999) Exp. Cell Res. 253, 1-6.

7.  White, M.F. et al. (1985) J. Biol. Chem. 260, 9470-9478.

8.  White, M.F. et al. (1988) J. Biol. Chem. 263, 2969-2980.

Entrez-Gene Id 3643
Swiss-Prot Acc. P06213

For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.
PathScan® is a trademark of Cell Signaling Technology, Inc.