Product Pathways - Tyrosine Kinase / Adaptors
Phospho-EGF Receptor Antibody Sampler Kit #9922
| Kit Includes | Quantity | Applications | Reactivity | MW (kDa) | Isotype |
|---|---|---|---|---|---|
| EGF Receptor (D38B1) XP® Rabbit mAb #4267 | 40 µl | W IP IHC-P IF-IC F | H M Mk | 175 | Rabbit IgG |
| Phospho-EGF Receptor (Tyr1068) (D7A5) XP® Rabbit mAb #3777 | 40 µl | W IHC-P IF-IC F | H M R Mk | 175 | Rabbit IgG |
| Phospho-EGF Receptor (Tyr992) Antibody #2235 | 40 µl | W IHC-P | H (R) | 175 | Rabbit |
| Phospho-EGF Receptor (Tyr1045) Antibody #2237 | 40 µl | W IF-IC | H R | 175 | Rabbit |
| Anti-rabbit IgG, HRP-linked Antibody #7074 | 100 µl | Goat |
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
Mk=Monkey
Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Specificity / Sensitivity
Each phospho-EGF Receptor antibody recognizes only the phosphorylated form of EGF Receptor at the indicated sites. The control EGF Receptor antibody recognizes both the phosphorylated and nonphosphorylated forms of EGF receptor.
Western Blotting
Western blot analysis of extracts from A431 cells, untreated or EGF-treated (100 ng/ml), using Phospho-EGF Receptor (Tyr992) Antibody #2235 (upper) or EGF Receptor Antibody #2232 (lower).
Western Blotting
Western blot analysis of extracts from A431 cells, untreated or EGF-stimulated (100 ng/ml), using Phospho-EGF Receptor (Tyr1045) Antibody #2237 (upper) or EGF Receptor (C74B9) Rabbit mAb #2646 (lower).
Western Blotting
Western blot analysis of extracts of BxPC-3 cells, untreated or EGF-stimulated, using Phospho-EGF Receptor (Tyr1068) (D7A5) Rabbit mAb #3777 (upper) and EGF Receptor Antibody #2232 (lower).
Description
The Phospho-EGF Receptor Antibody Sampler Kit provides an economical means of evaluating the EGF Receptor and several phosphorylation sites that are involved in its activation. The kit contains enough primary and secondary antibodies to perform four Western blot experiments.
Source / Purification
Antibodies are produced by immunizing animals with synthetic phosphopeptides corresponding to residues surrounding Tyr992, Tyr1045 or Tyr1068 of human EGF receptor. EGF Receptor Antibody is produced by immunizing animals with a fusion protein containing the cytoplasmic domain of human EGF receptor. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.
Background
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).
- Hackel, P.O. et al. (1999) Curr Opin Cell Biol 11, 184-9.
- Zwick, E. et al. (1999) Trends Pharmacol Sci 20, 408-12.
- Cooper, J.A. and Howell, B. (1993) Cell 73, 1051-4.
- Hubbard, S.R. et al. (1994) Nature 372, 746-54.
- Biscardi, J.S. et al. (1999) J Biol Chem 274, 8335-43.
- Emlet, D.R. et al. (1997) J Biol Chem 272, 4079-86.
- Levkowitz, G. et al. (1999) Mol Cell 4, 1029-40.
- Ettenberg, S.A. et al. (1999) Oncogene 18, 1855-66.
- Rojas, M. et al. (1996) J Biol Chem 271, 27456-61.
- Feinmesser, R.L. et al. (1999) J Biol Chem 274, 16168-73.
Application References
- Wang, X. et al. (2011) Blood 118, 1255-63. Applications: Western Blotting
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Protocols
- 2235:
- IHC / Paraffin, Western Blotting
- 2237:
- Immunofluorescence, Western Blotting
- 3777:
- Flow, IHC / Paraffin, Immunofluorescence, Western Blotting
- 4267:
- Flow, IHC / Paraffin, Immunofluorescence, Immunoprecipitation, Western Blotting
Companion Products
- 9923 Phospho-HER2/ErbB2 Antibody Sampler Kit
- 7071 Phototope®-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 7727 Biotinylated Protein Ladder Detection Pack
- 7003 20X LumiGLO® Reagent and 20X Peroxide
For Research Use Only. Not For Use In Diagnostic Procedures.