Cell Signaling Technology

Product Pathways - Glucose Metabolism

Phospho-Insulin Receptor β (Tyr1361) (84B2) Rabbit mAb #3023

Applications Reactivity Sensitivity MW (kDa) Isotype
W H Transfected Only 95 KDa Rabbit IgG

Applications Key:  W=Western Blotting
Reactivity Key:  H=Human
Species cross-reactivity is determined by Western blot.

Specificity / Sensitivity

Phospho-Insulin Receptor β (Tyr1361) (84B2) Rabbit mAb detects transfected levels of insulin receptor β only when phosphorylated at Tyr1361. It slightly cross-reacts with activated IGF-I receptor, but does not cross-react with other activated tyrosine kinases.

Source / Purification

Monoclonal antibodies are produced by immunizing rabbits with a synthetic phosphopeptide (KLH-coupled) corresponding to residues surrounding Tyr1361 of human insulin receptor β.

Western Blotting

Western Blotting

Cross-reactivity testing of Phospho-Insulin Receptor (Tyr1361) (84B2) Rabbit mAb with activated IGF-I receptors. Various amounts of activated recombinant IGF-I receptor cytoplasmic domain proteins were applied for Western blot analysis, using Phospho-Insulin Receptor (Tyr1361) (84B2) Rabbit mAb (upper) and Phospho-IGF-I Receptor (Tyr1315/1316) (19H7) Rabbit mAb #3024 (lower) respectively. The result showed that Phospho-Insulin Receptor (Tyr1361) (84B2) Rabbit mAb barely cross-reacts with activated IGF-I receptors.

Western Blotting

Western Blotting

Western blot analysis of CHO cells overexpressing human insulin receptors untreated or treated with insulin (100 nM for 5 min), using Phospho-Insulin Receptor β (Tyr1361) (84B2) Rabbit mAb (upper) and Insulin Receptor β (4B8) Rabbit mAb #3025(lower).

IF-IC

IF-IC

Confocal immunofluorescent analysis of CHO cells overexpressing human insulin receptors, unstimulated (left) and stimulated (right) with insulin, using Phospho-Insulin Receptor β (Tyr1361) (84B2) Rabbit mAb (red).


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 insulin receptor is one of the earliest cellular responses to insulin stimulation (7). Autophosphorylation begins with phosphorylation of Tyr1146 and either Tyr1150 or Tyr1151, while full kinase activation requires the 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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Companion Products

Rabbit Monoclonals Produced Using Epitomics® Technology, U.S. Patent No. 5,675,063.

This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.

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