Cell Signaling Technology

Product Pathways - Tyrosine Kinase/ Adaptors

Phospho-EGF Receptor (Tyr1068) (1H12) Mouse mAb #2236

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP IHC-P IF-IC F H M R Endogenous 175 Mouse IgG1

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat
Species cross-reactivity is determined by Western blot.

Specificity / Sensitivity

Phospho-EGF Receptor (Tyr1068) (1H12) Mouse mAb detects endogenous levels of EGF receptor only when phosphorylated at Tyr1068. This antibody does not recognize EGF receptor phosphorylated at other sites, but may cross-react with other activated ErbB family members.

Source / Purification

Monoclonal antibody is produced by immunizing mice with a synthetic phospho-peptide (KLH-coupled) corresponding to residues surrounding Tyr1068 of human EGF receptor.

Western Blotting

Western Blotting

Western blot analysis of extracts from A431 cells, untreated or EGF-stimulated for various times, using Phospho-EGF Receptor (Tyr1068) (1H12) Mouse mAb (upper) or EGF Receptor Antibody #2232 (lower).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, showing membrane localization, using Phospho-EGF Receptor (Tyr1068) (1H12) Mouse mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, untreated (left) or calf intestinal phosphatase (CIP) treated (right), using Phospho-EGF Receptor (Tyr1068) (1H12 ) Mouse mAb.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analyis of paraffin-embedded human glioma using Phospho-EGF Receptor (Tyr1068) (1H12) Mouse mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded MDA-MB-468 cells, untreated (left) or EGF-treated (right), using Phospho-EGF Receptor (Tyr1068) (1H12) Mouse mAb.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of A431 cells, untreated (left) or EGF-treated (right), using Phospho-EGF Receptor (Tyr1068) 1H12 Mouse mAb and EGF Receptor Antibody #2232.


IF-P

IF-P

Confocal immunofluorescent analysis of paraffin embedded human lung squamous cell carcinoma using Phospho-EGF Receptor (Tyr1068) (1H12) Mouse mAb (red). Blue pseudocolor = DRAQ5™ (fluorescent DNA dye).

Background

The epidermal growth factor (EGF) receptor is a 170 kDa transmembrane tyrosine kinase that belongs to the HER/ErbB protein family. Ligand binding results in receptor dimerization, autophosphorylation, activation of downstream signaling and lysosomal degradation (1,2). Phosphorylation of EGF receptor (EGFR) at Tyr845 in the kinase domain is implicated in stabilizing the activation loop, maintaining the active state enzyme and providing a binding surface for substrate proteins (3,4). c-Src is involved in phosphorylation of EGFR at Tyr845 (5). The SH2 domain of PLCγ binds at phospho-Tyr992, resulting in activation of PLCγ-mediated downstream signaling (6). Phosphorylation of EGFR at Tyr1045 creates a major docking site for c-Cbl, an adaptor protein that leads to receptor ubiquitination and degradation following EGFR activation (7,8). The GRB2 adaptor protein binds activated EGFR at phospho-Tyr1068 (9). A pair of phosphorylated residues (Tyr1148 and Tyr1173) provides a docking site for the SHC scaffold protein, with both sites involved in MAP kinase signaling activation (2). Phosphorylation of EGFR at specific serine and threonine residues attenuates EGFR kinase activity. EGFR carboxy-terminal residues Ser1046 and Ser1047 are phosphorylated by CaM kinase II; mutations to either of these serines results in upregulated EGFR tyrosine autophosphorylation (10).

  1. Hackel, P.O. et al. (1999) Curr. Opin. Cell Biol. 11, 184-189.
  2. Zwick, E. et al. (1999) Trends Pharmacol. Sci. 20, 408-412.
  3. Cooper, J.A. and Howell, B. (1993) Cell 73, 1051-1054.
  4. Hubbard, S.R. et al. (1994) Nature 372, 746-754.
  5. Biscardi, J.S. et al. (1999) J. Biol. Chem. 274, 8335-8343.
  6. Emlet, D.R. et al. (1997) J. Biol. Chem. 272, 4079-4086.
  7. Levkowitz, G. et al. (1999) Mol. Cell 4, 1029-1040.
  8. Ettenberg, S.A. et al. (1999) Oncogene 18, 1855-1866.
  9. Rojas, M. et al. (1996) J. Biol. Chem. 271, 27456-27461.
  10. Feinmesser, R.L. et al. (1999) J. Biol. Chem. 274, 16168-16173.

Application References

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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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