EGFR [p Tyr1197] Antibody - BSA Free

Novus Biologicals | Catalog # NBP1-77936

Novus Biologicals
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Key Product Details

Species Reactivity

Human

Applications

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, ELISA

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all EGFR Primary Antibodies »

Product Specifications

Immunogen

This affinity purified EGFR [p Tyr1197] Antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding to the C-Terminus near amino acids 1175-1200 of human EGFR protein. (Uniprot: P00533)

Reactivity Notes

A BLAST analysis was used to suggest cross reactivity with phosphorylated EGFR from human, mouse and rat sources. Reactivity of this antibody with EGFR from other species is unknown.

Modification

p Tyr1197

Specificity

This affinity-purified antibody is directed against the phosphorylated form of human EGFR protein at the pY1197 residue. The resultant affinity purified antibody was then cross-adsorbed against the non-phosphorylated form of the immunizing peptide. Reactivity occurs against human EGFR pY1197 protein and the antibody is specific for the phosphorylated form of the protein. Reactivity with non-phosphorylated human EGFR is minimal by ELISA. A BLAST analysis was used to suggest cross reactivity with phosphorylated EGFR from human, mouse and rat sources. Reactivity of this antibody with EGFR from other species is unknown.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Description

This affinity-purified antibody is directed against the phosphorylated form of human EGFR protein at the [p Tyr1197] residue. The product was affinity purified from monospecific antiserum by immunoaffinity purification. Antiserum was first purified against the phosphorylated form of the immunizing peptide. The resultant affinity purified antibody was then cross-adsorbed against the non-phosphorylated form of the immunizing peptide

Store vial at -20C prior to opening. Aliquot contents and freeze at -20C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4C as an undiluted liquid. Dilute only prior to immediate use.

Scientific Data Images for EGFR [p Tyr1197] Antibody - BSA Free

Western Blot: EGFR [p Tyr1197] Antibody [NBP1-77936]

Western Blot: EGFR [p Tyr1197] Antibody [NBP1-77936]

Western Blot: EGF R/ErbB1 [p Tyr1197] Antibody [NBP1-77936] - Lane 1: unstimulated A431 cells. Lane 2: A431 cells stimulated with EGF (50 ng/ml for 15 min). Load: 30 ug per lane. Primary antibody: EGFR pY1197 antibody at 1:250 for overnight at 4C. Secondary antibody: IRDye800 (TM) rabbit secondary antibody at 1:10,000 for 45 min at RT. Block: 5% BLOTTO overnight at 4C. Predicted/Observed size: 134.3 kDa, ~170 kDa for EGFR pY1197.
Immunohistochemistry-Paraffin: EGFR [p Tyr1197] Antibody [NBP1-77936]

Immunohistochemistry-Paraffin: EGFR [p Tyr1197] Antibody [NBP1-77936]

Immunohistochemistry-Paraffin: EGF R/ErbB1 [p Tyr1197] Antibody [NBP1-77936] - Tissue: placental trophoblasts. Fixation: formalin fixed paraffin embedded. Antigen retrieval: not required. Primary antibody: EGFR pY1197 antibody at 5ug/mL for 1 h at RT. Secondary antibody: Peroxidase rabbit secondary antibody at 1:10,000 for 45 min at RT. Localization: EGFR pY1197 is on the cell membrane. Staining: EGFR pY1197 as precipitated red signal with hematoxylin purple counterstain.
EGFR [p Tyr1197] Antibody

EGFR [p Tyr1197] Antibody

Immunohistochemistry of Rabbit anti-EGFR pY1197 antibody. Tissue: placental trophoblasts. Fixation: formalin fixed paraffin embedded. Antigen retrieval: not required. Primary antibody: EGFR pY1197 antibody at 5 ug/ml for 1 h at RT. Secondary antibody: Peroxidase rabbit secondary antibody at 1:10,000 for 45 min at RT. Localization: EGFR pY1197 is on the cell membrane. Staining: EGFR pY1197 as precipitated red signal with hematoxylin purple counterstain.
EGFR [p Tyr1197] Antibody

EGFR [p Tyr1197] Antibody

Western Blot of Rabbit anti-EGFR pY1197 antibody. Lane 1: unstimulated A431 cells

Applications for EGFR [p Tyr1197] Antibody - BSA Free

Application
Recommended Usage

ELISA

1:4000-1:20000

Immunohistochemistry

5 ug/ml

Immunohistochemistry-Paraffin

1:10-1:500

Western Blot

1:250-1:1500
Application Notes
This affinity purified antibody has been tested for use in ELISA, immunohistochemistry and western blot. Specific conditions for reactivity should be optimized by the end user. Expect a band approximately 170 kDa in size corresponding to phosphorylated EGFR protein by western blotting in the appropriate cell lysate or extract. Less than 5.0% reactivity is observed against the non-phosphorylated form of the immunizing peptide. This antibody is phospho specific for pY1197 of EGFR protein.

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2

Format

BSA Free

Preservative

0.01% Sodium Azide

Concentration

Please see the vial label for concentration. If unlisted please contact technical services.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at -20C short term. Aliquot and store at -80C long term. Avoid freeze-thaw cycles.

Background: EGFR

Epidermal growth factor receptor (EGFR), also known as ErbB1 and HER1, is a type I glycoprotein that belongs the ErbB subfamily of receptor tyrosine kinases (RTKs), which includes ErbB2/HER2, ErbB3/HER3, and ErbB4/HER4 (1,2). EGFR plays an important role in epithelial cell development and homeostasis and as a driver of tumorigenesis in cancer (1,2). The human EGFR is protein 1210 amino acids (aa) in length with a theoretical molecular weight (MW) of 134 kDa (1). The protein consists of a short signal peptide, an extracellular domain (ECD) divided into four subdomains (I-IV), a transmembrane region, an intracellular juxtamembrane segment, a tyrosine kinase domain, and C-terminal tail (1-3). Within the ECD, human EGFR has 88-90% aa sequence identity with mouse and rat EGFR. EGFR has four known specific ligands: EGF, amphiregulin, epigen, and transforming growth factor alpha (TGF-alpha). EGFR ligands betacellulin, epiregulin, and herapin binding (HB)-EGF have dual specificity with ErbB4 (1,3). Ligand binding to the extracellular domain of EFGR leads to receptor homodimerization, or heterodimerization with other ErbB family members, and EGFR activation. This results in subsequent phosphorylation and activation of intracellular signaling pathways, such as MAPK and PI3K/Akt (2,3). EGFR signaling is essential for many cellular processes including proliferation, differentiation, migration, and apoptosis (1,3,5).

In addition to its role in normal development, EGFR mutations or overexpression is observed in many tumors, including breast cancer, non-small cell lung carcinoma (NSCLC), colon cancer, and more (3-6). Small molecule tyrosine kinase inhibitors (TKIs), like gefitinib, erlotinib, and afatinib, have shown great efficacy in treating patients with EGFR activating mutations, especially for NSCLC (4-6). However, most patients eventually develop acquired resistance to TKIs and thus combination and alternative therapies are in development (4-6). A third-generation TKI, osimertinib, is approved for NSCLC patients with resistance to first-line EGFR TKI treatment (6). Additionally, combination therapies of EGFR TKIs with monoclonal antibody immunotherapies, like anti-PD-L1, are being further investigated in clinical trials (6).

References

1. Roskoski R Jr. Small molecule inhibitors targeting the EGFR/ErbB family of protein-tyrosine kinases in human cancers. Pharmacol Res. 2019; 139:395-411. https://doi.org/10.1016/j.phrs.2018.11.014

2. Sigismund S, Avanzato D, Lanzetti L. Emerging functions of the EGFR in cancer. Mol Oncol. 2018; 12(1):3-20. https://doi.org/10.1002/1878-0261.12155

3. Normanno N, De Luca A, Bianco C, et al. Epidermal growth factor receptor (EGFR) signaling in cancer. Gene. 2006; 366(1):2-16. https://doi.org/10.1016/j.gene.2005.10.018

4. Liu Q, Yu S, Zhao W, Qin S, Chu Q, Wu K. EGFR-TKIs resistance via EGFR-independent signaling pathways. Mol Cancer. 2018; 17(1):53. https://doi.org/10.1186/s12943-018-0793-1

5. Harrison PT, Vyse S, Huang PH. Rare epidermal growth factor receptor (EGFR) mutations in non-small cell lung cancer. Semin Cancer Biol. 2020; 61:167-179. https://doi.org/10.1016/j.semcancer.2019.09.015

6. Wu SG, Shih JY. Management of acquired resistance to EGFR TKI-targeted therapy in advanced non-small cell lung cancer. Mol Cancer. 2018; 17(1):38. https://doi.org/10.1186/s12943-018-0777-1

Long Name

Epidermal Growth Factor Receptor

Alternate Names

EGF R, ErbB, ErbB1, HER-1

Gene Symbol

EGFR

UniProt

P00533, NP_005219 (Human)

Additional EGFR Products

Product Documents for EGFR [p Tyr1197] Antibody - BSA Free

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Product Specific Notices for EGFR [p Tyr1197] Antibody - BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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Protocols

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