Product Pathways - Tyrosine Kinase/ Adaptors
EGF Receptor Blocking Peptide (IHC Preferred) #1165
Description
This peptide can be used to block EGF Receptor (15F8) Rabbit mAb (IHC Preferred) #4405.
Quality Control
The quality of the peptide was evaluated by reversed-phase HPLC and by mass spectrometry. The peptide blocks EGF Receptor (15F8) Rabbit mAb (IHC Specific) #4405 signal completely in immunohistochemistry.
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using EGF Receptor (15F8) Rabbit mAb (IHC Preferred) preincubated with an irrelevant control peptide (left) or the same antibody preincubated with EGF Receptor Blocking Peptide (IHC Preferred) (right).
Applications
Use as a blocking reagent to evaluate the specificity of antibody reactivity in immunohistochemistry protocols.
Directions for Use
For Immunohistochemistry, add twice the volume of peptide as volume of antibody used in a 100 ul total volume. Incubate for a minimum of 30 minutes prior to adding the entire volume to the slide. Recommended antibody dilutions can be found on the EGF Receptor (15F8) Rabbit mAb (IHC Preferred) #4405 datasheet.
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).
- Hackel, P.O. et al. (1999) Curr. Opin. Cell Biol. 11, 184-189.
- Zwick, E. et al. (1999) Trends Pharmacol. Sci. 20, 408-412.
- Cooper, J.A. and Howell, B. (1993) Cell 73, 1051-1054.
- Hubbard, S.R. et al. (1994) Nature 372, 746-754.
- Biscardi, J.S. et al. (1999) J. Biol. Chem. 274, 8335-8343.
- Emlet, D.R. et al. (1997) J. Biol. Chem. 272, 4079-4086.
- Levkowitz, G. et al. (1999) Mol. Cell 4, 1029-1040.
- Ettenberg, S.A. et al. (1999) Oncogene 18, 1855-1866.
- Rojas, M. et al. (1996) J. Biol. Chem. 271, 27456-27461.
- Feinmesser, R.L. et al. (1999) J. Biol. Chem. 274, 16168-16173.
Application References
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
Companion Products
- 4405 EGF Receptor (15F8) Rabbit mAb (IHC Preferred)
- 8102 SignalSlide™ Phospho-EGF Receptor (Tyr1173) IHC Controls
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.
