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Primary Antibodies
EGFR [pY1068] Antibody
Background
Epidermal growth factor receptor (EGFR, a 185 kDa glycoprotein), also known as ErbB-1, is a transmembrane tyrosine kinase that regulates a variety of biological responses ranging from mitogenesis to stress signaling. The EGFR consists of a large extracellular domain, a single transmembrane domain and a cytoplasmic domain that exhibits kinase activity. Upon binding of EGF to the extracellular domain, the receptor undergoes dimerization and becomes phosphorylated on several tyrosine residues within the cytoplasmic domain. These events result in EGFR activation and increased tyrosine kinase activity toward a variety of intracellular substrates. Tyrosine 1068, within the cytoplasmic domain of the receptor, is a major autophosphorylation site that allows binding of Grb2 and activation of the Ras → Raf → ERK1&2 signaling pathway.
![EGFR [pY1068] Antibody](http://www.prosci-inc.com/shop/images/XBP-4086.gif)
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Description
Lysates from A431 cells unstimulated (lane 1) or stimulated with 200 ng/mL EGF for 15 minutes (lanes 2-5), were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to PVDF. The membrane was blocked with a 5% BSA-TBST buffer for one hour at room temperature, and then incubated with the EGFR [pY1068] antibody for two hours at room temperature in a 1% BSA-TBST buffer, following prior incubation with: no peptide (lanes 1 and 2), the non-phosphopeptide corresponding to the phosphopeptide immunogen (lane 3), a generic phosphotyrosine-containing peptide (lane 4) or the phosphopeptide immunogen (lane 5). After washing, the membrane was incubated with goat F(ab’)2 anti-rabbit IgG HRP conjugate and signals were detected using the Pierce SuperSignal™ method.
The data show that only the phosphopeptide corresponding to EGFR [pY1068] blocks the antibody signal, demonstrating the specificity of the antibody. The data also show the induction of EGFR [pY1068] phosphorylation by the addition of EGF to this cell system.
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Source
EGFR antibody was produced against a chemically synthesized phosphopeptide derived from the region of human EGFR that contains tyrosine 1068. The sequence is conserved in rat.
Purification
EGFR antibody was purified from rabbit serum by sequential epitope-specific chromatography. The antibody has been negatively preadsorbed using a non-phosphopeptide corresponding to the site of phosphorylation to remove antibody that is reactive with non-phosphorylated epidermal growth factor receptor (EGFR). The final product is generated by affinity chromatography using an EGFR-derived peptide that is phosphorylated at tyrosine 1068.
Clonality / Clone
This is a polyclonal antibody.
Host
EGFR [pY1068] antibody was raised in rabbit.
Please use anti-rabbit secondary antibodies.
Specificity
Human EGFR. Mouse and rat (100% homologous) EGFR have not been tested, but are expected to react.
Application
For Western blotting applications, we recommend using the antibody at a 1:1000 dilution.
Tested Application
WB
Buffer
Rabbit polyclonal immunoglobulin in Dulbecco’s phosphate buffered saline (without Mg2+ and Ca2+), pH 7.3 (+/- 0.1), 50% glycerol with 1.0 mg/mL BSA (IgG, protease free) as a carrier.
Storage
Store at -20˚C. We recommend a brief centrifugation before opening to settle vial contents. Then, apportion into working aliquots and store at -20˚C. For shipment or short-term storage (up to one week), 2-8˚C is sufficient.
Positive Control
- Human epidermoid carcinoma (A431) cells +/- EGF.
Species Reactivity
H, M, R
Formulation
Rabbit polyclonal immunoglobulin in Dulbecco’s phosphate buffered saline (without Mg2+ and Ca2+), pH 7.3 (+/- 0.1), 50% glycerol with 1.0 mg/mL BSA (IgG, protease free) as a carrier.
Short Description
EGFR [pY1068]
This product belongs to the following categories:
References- Cuadrado, A., et al. (2003) Aplidin induces apoptosis in human cancer cells via glutathione depletion and sustained activation of the epidermal growth factor receptor, Src, JNK, and p38 MAPK. J. Biol. Chem. 278(1):241-250.
- Kalyankrishna, S. and K.U. Malik (2003) Norepinephrine-induced stimulation of p38 mitogen-activated protein kinase is mediated by arachidonic acid metabolites generated by activation of cytosolic phospholipase A(2) in vascular smooth muscle cells. J. Pharmacol. Exp. Ther. 304(2):761-772.
- Moro, L., et al. (2002) Integrin-induced epidermal growth factor (EGF) receptor activation requires c-Src and p130Cas and leads to phosphorylation of specific EGF receptor tyrosines. J. Biol. Chem. 277(11):9405-9414.
Datasheet 0801-W
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