Western blot analysis of extracts from A431 cells, untreated or EGF-stimulated, using Phospho-EGF Receptor (Tyr1068) Antibody #2234 (left) or the same antibody preincubated with Phospho-EGF Receptor (Tyr1068) Antibody Blocking Peptide (right).Learn more about how we get our images
This peptide is used to block Phospho-EGF Receptor (Tyr1068) Antibody #2234 and Phospho-EGF Receptor (Tyr1068) (D7A5) XP® Rabbit mAb #3777 reactivity.
The quality of the peptide was evaluated by reversed-phase HPLC and by mass spectrometry. The peptide blocks Phospho-EGF Receptor (Tyr1068) Antibody #2234 and Phospho-EGF Receptor (Tyr1068) (D7A5) XP® Rabbit mAb #3777 signal completely in western blotting and immunohistochemistry.
Use as a blocking reagent to evaluate the specificity of antibody reactivity in western immunoblotting and Immunohistochemistry protocols. For western Immunoblotting, add 10 ul of antibody and 10 ul of blocking peptide to 10 ml of antibody dilution buffer, and incubate at room temperature for 1 hour before allowing to react with the blot.
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 Phospho-EGF receptor (Tyr1068) Antibody #2234 and Phospho-EGF Receptor (Tyr1068) (D7A5) XP® Rabbit mAb #3777 data sheets.
Supplied in 20 mM potassium phosphate (pH 7.0), 50 mM NaCl, 0.1 mM EDTA, 1 mg/ml BSA, 5% glycerol and 1% DMSO. Store at –20°C.
The epidermal growth factor (EGF) receptor is a transmembrane tyrosine kinase that belongs to the HER/ErbB protein family. Ligand binding results in receptor dimerization, autophosphorylation, activation of downstream signaling, internalization, 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 the adaptor protein c-Cbl, leading 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 EGFR residues (Tyr1148 and Tyr1173) provide 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; mutation of either of these serines results in upregulated EGFR tyrosine autophosphorylation (10).
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