Cell Signaling Technology

Product Pathways - PathScan ELISA

PathScan® Phospho-IGF-I Receptor β (Tyr1131) Sandwich ELISA Antibody Pair #7820

Kit Includes Volume Cap Color
P-IGF-I Receptor beta(Y1131) Capture Ab (100X) 0.4 ml Pink
IGF-I Receptor Detection Ab (100X) 0.4 ml Blue
Anti-mouse IgG, HRP-linked Antibody (1000X) 0.04 ml Yellow

Capture and detection antibodies are stored at 4°C. HRP-linked secondary reagent is stored at -20°C.

Species Cross-Reactivity

H

Reactivity Key:  H=Human
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Description

CST's PathScan® Phospho-IGF-I Receptor β (Tyr1131) Sandwich ELISA Antibody Pair is offered as an economical alternative to our PathScan® Phospho-IGF-I Receptor β (Tyr1131) Sandwich ELISA Kit #7302. Capture and detection antibodies (100X stocks) and HRP-linked secondary antibody (1000X stock) are supplied. Sufficient reagents are supplied for 4 x 96 well ELISAs. The phospho-IGF-I receptor β (Tyr1131) capture antibody is coated on a 96 well microplate in PBS overnight. After blocking, cell lysates are added followed by an IGF-I receptor detection antibody and anti-mouse IgG, HRP-linked antibody. HRP substrate, TMB, is added for color development. The magnitude of the absorbance for this developed color is proportional to the quantity of phospho-IGF-I receptor β (Tyr1131) protein.Antibodies in kit are custom formulations specific to kit.

Specificity / Sensitivity

For Antibody Pair specificity and sensitivity, please refer to the corresponding PathScan® Sandwich ELISA Kit. Note: This antibody pair detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

ELISA

ELISA

The relationship between protein concentration of lysates from untreated or IGF-I-treated MCF-7 cells and the absorbance at 450 nm using PathScan® Phospho-IGF-I Receptor β (Tyr1131) Sandwich ELISA Antibody Pair is shown. Cells were serum starved overnight, treated with 100 nm IGF-I at 37°C for 5 minutes, and then lysed.

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).

  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.

Application References

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For Research Use Only. Not For Use In Diagnostic Procedures.

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