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General description
Epidermal growth factor receptor (UniProt: P00533; also known as EC:2.7.10.1, Proto-oncogene c-ErbB-1, Receptor tyrosine-protein kinase erbB-1) is encoded by the EGFR (also known as ERBB, ERBB1, HER1) gene (Gene ID: 1956) in human. EGFR is a single-pass type I membrane glycoprotein that is ubiquitously expressed. It is synthesized with a signal peptide (1-24), which is subsequently cleaved off to produce the mature form that contains an extracellular domain (aa 25-645), a transmembrane domain (aa 646-668), and a cytoplasmic domain (669-1210). It possesses receptor tyrosine kinase activity and upon binding of ligands activates several signaling cascades. Ligand binding is known to trigger receptor homo- and/or heterodimerization, which stimulates its intrinsic intracellular protein-tyrosine kinase activity. As a result, autophosphorylation of tyrosine residues in the C-terminal domain of EGFR occurs, which results in activation of several downstream signaling cascades, including the Ras-RAF-MEK-ERK, PI3 kinase-Akt, PLCγ-PKC, and STAT. EGFR phosphorylated on at serine 695 is reported to be only partial and occurs only if threonine 693 is phosphorylated. Tyrosine 1068 is one of the major autophosphorylation sites. Phosphorylation at tyrosine 1068 is considered as a powerful biomarker associated with erlotinib sensitivity in vitro and in preclinical cancer stem cells generated xenografts. EGFR also undergoes mono and polyubiquitination upon EGF stimulation. However, this ubiquitination does not affect tyrosine kinase activity of the receptor or its signaling capacity. EGFR is commonly overexpressed and is mutated in many human malignancies and is often associated aggressive phenotypes. This ZooMAb® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Sette, G., et al. (2015). Cell Death Dis. 6(8); e1850; Wang, F., et al. (2012). J. Exp. Clin. Cancer Res. 31; Article 65).
We are committed to bringing you greener alternative products, which adhere to one or more of The 12 Principles of Green Chemistry. This antibody is Preservative-free, produced without the harm or sacrifice of animals and exceptionally stable to allow for ambient shipping and storage if needed and thus aligns with "Waste Prevention", "Designing Safer Chemicals" and "Design for Energy Efficiency". Click here for more information.
ZooMAb® antibodies represent an entirely new generation of recombinant monoclonal antibodies. Each ZooMAb® antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb® antibodies are reliably available and ready to ship when you need them.
Immunogen
KLH-conjugated linear peptide corresponding to 12 amino acids surrounding phosphotyrosine 1068 from the C-terminal region of human Epidermal growth factor receptor (EGFR).
Application
Quality Control Testing
Evaluated by Western Blotting in lysate from A431 cells treated with EGF.
Western Blotting Analysis (WB): A 1:10,000 dilution of this antibody detected EGFR phosphorylated on Tyrosine 1068 in lysate from over-night serum starved A431 cells treated with EGF (50 ng/mL; 10 min.).
Tested Applications
Western Blotting Analysis: A 1:10,000 dilution from a representative lot detected Phospho-EGFR (Tyr1068) in lysate from HeLa cells overnight serum starved and treated with EGF ( (50 ng/mL; 10 min.).
Affinity Binding Assay: A representative lot of this antibody bound Phospho-EGF Receptor-(Tyr1068) peptide with a KD of 1.1 x 10-6 in an affinity binding assay.
Flow Cytometry Analysis: 1 µg from a representative lot detected Phospho-EGFR (Tyr1068) in one million A431 cells treated with EGF (100 ng/mL; 15 min.).
Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected Phospho-EGFR (Tyr1068) in human cervical cancer tissue sections.
Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected Phospho-EGFR (Tyr1068) in A431 cells treated with EGF (200 ng/mL; 10 min.).
Peptide Inhibition Assay: Target band detection in a lysate from A431 cells treated with EGF (50 ng/mL; 10 min) was prevented by preblocking of a representative lot with the immunogen phosphopeptide, but not the corresponding non-phosphopeptide.
Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user.
Biochem/physiol Actions
Clone 3F4 is a ZooMAb® rabbit recombinant monoclonal antibody that specifically detects Epidermal growth factor receptor (EGFR) phosphorylated on tyrosine 1068.
Physical form
Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS, 5% Trehalose, normal appearance a coarse or translucent resin. The PBS/trehalose components in the ZooMAb formulation can have the appearance of a semi-solid (bead like gel) after lyophilization. This is a normal phenomenon. Please follow the recommended reconstitution procedure in the data sheet to dissolve the semi-solid, bead-like, gel-appearing material. The resulting antibody solution is completely stable and functional as proven by full functional testing. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 µL.
Preparation Note
Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 µL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws.
Reconstitute lyophilized antibody pellet with 25μL of ultrapure water or Phosphate Buffered Saline (PBS). Please refer to our reconstitution protocol and the specific application guidance on the suggested starting dilutions and sample type.
Other Notes
Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.
Legal Information
ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany
Disclaimer
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
| biological source | rabbit |
| Quality Segment | 200 |
| recombinant | expressed in HEK 293 cells |
| conjugate | unconjugated |
| antibody form | purified antibody |
| antibody product type | primary antibodies |
| clone | 3F4, recombinant monoclonal |
| description | 3F4 Clone |
| product line | ZooMAb® learn more |
| form | lyophilized |
| mol wt | calculated mol wt 134.28 kDa,observed mol wt ~160 kDa |
| purified by | using Protein A |
| species reactivity | human |
| species reactivity (predicted by homology) | monkey |
| packaging | antibody small pack of 25 μL |
| greener alternative product characteristics | Waste Prevention Designing Safer Chemicals Design for Energy Efficiency Learn more about the Principles of Green Chemistry. |
| enhanced validation | recombinant expression Learn more about Antibody Enhanced Validation |
| technique(s) | affinity binding assay: suitable,flow cytometry: suitable,immunocytochemistry: suitable,immunohistochemistry: suitable,western blot: suitable |
| isotype | IgG |
| epitope sequence | C-terminal |
| Protein ID accession no. | NP_005219 |
| UniProt accession no. | P00533 |
| greener alternative category | Aligned |
| shipped in | ambient |
| storage temp. | 2-8°C |
| Gene Information | human ... EGFR(1956) |
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