Product Pathways - PathScan ELISA
PathScan® Phospho-Insulin Receptor β (Tyr1150/1151) Sandwich ELISA Kit #7258
|7258C||1 Kit (96 assays)||---||In Stock||---|
|7258||carrier free and custom formulation / quantity||email request|
When ordering five or more kits, please contact us for processing time and pricing at firstname.lastname@example.org.
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|Kit 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|
Note: 12 8-well modules – Each module is designed to break apart for 8 tests.
Storage: Kit should be stored at 4°C with the exception of Cell Lysis Buffer, which is stored at –20°C (packaged separately).
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.
ELISA - Western correlation
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.
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).
- Adams, T.E. et al. (2000) Cell. Mol. Life Sci. 57, 1050-1093.
- Baserga, R. et al. (2000) Oncogene 19, 5574-5581.
- Scheidegger, K.J. et al. (2000) J. Biol. Chem. 275, 38921-38928.
- Hernandez-Sanchez, C. et al. (1995) J. Biol. Chem. 270, 29176-29181.
- Lopaczynski, W. et al. (2000) Biochem. Biophys. Res. Commun. 279, 955-960.
- Baserga, R. et al. (1999) Exp. Cell Res. 253, 1-6.
- White, M.F. et al. (1985) J. Biol. Chem. 260, 9470-9478.
- White, M.F. et al. (1988) J. Biol. Chem. 263, 2969-2980.
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- 7082 PathScan® Phospho-Insulin Receptor β (panTyr) Sandwich ELISA Kit
- 7069 PathScan® Total Insulin Receptor β Sandwich ELISA Kit
- 3024 Phospho-IGF-I Receptor β (Tyr1135/1136)/Insulin Receptor β (Tyr1150/1151) (19H7) Rabbit mAb
- 3025 Insulin Receptor β (4B8) Rabbit mAb
- 3020 Insulin Receptor β (L55B10) Mouse mAb
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 9803 Cell Lysis Buffer (10X)
- 7004 TMB Substrate
- 7002 STOP Solution
For Research Use Only. Not For Use In Diagnostic Procedures.
PathScan® is a trademark of Cell Signaling Technology, Inc.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.