EGFR [p Tyr1045] Antibody (PSH03-28)

Novus Biologicals, part of Bio-Techne | Catalog # NBP3-32760

Recombinant Monoclonal Antibody
Novus Biologicals, part of Bio-Techne
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NBP3-32760
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Key Product Details

Species Reactivity

Human

Applications

Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Western Blot

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Clone # PSH03-28

Concentration

1 mg/ml

View all EGFR Antibodies »

Product Specifications

Immunogen

Synthetic phosphopeptide corresponding to residues surrounding Tyr1045 of EGFR. (Uniprot: P00533)

Modification

p Tyr 1045

Localization

Cell membrane, Nucleus membrane, Nucleus, Endoplasmic reticulum membrane, Golgi apparatus membrane, Endosome.

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

134 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for EGFR [p Tyr1045] Antibody (PSH03-28)

EGFR [p Tyr1045] Antibody (PSH03-28)

Western Blot: EGFR [p Tyr1045] Antibody (PSH03-28) [NBP3-32760]

Western Blot: EGFR [p Tyr1045] Antibody (PSH03-28) [NBP3-32760] - Western blot analysis of EGFR on different lysates with Rabbit anti-EGFR antibody (NBP3-32760) at 1/1,000 dilution.

Lane 1: A431 cell lysate
Lane 2: A431 treated with 100ng/mL EGF for 30 minutes cell lysate

Lysates/proteins at 30 ug/Lane.

Predicted band size: 134 kDa
Observed band size: 170 kDa

Exposure time: 3 minutes 10 seconds;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (NBP3-32760) at 1/1,000 dilution was used in 5% NFDM/TBST at 4C overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
EGFR [p Tyr1045] Antibody (PSH03-28)

Immunocytochemistry/ Immunofluorescence: EGFR [p Tyr1045] Antibody (PSH03-28) [NBP3-32760]

Immunocytochemistry/ Immunofluorescence: EGFR [p Tyr1045] Antibody (PSH03-28) [NBP3-32760] - Immunocytochemistry analysis of A431 treated with or without 100ng/mL EGF for 60 minutes cells labeling EGFR with Rabbit anti-EGFR antibody (NBP3-32760) at 1/100 dilution.

Cells were fixed in 100% precooled methanol for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-EGFR antibody (NBP3-32760) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.
EGFR [p Tyr1045] Antibody (PSH03-28)

Flow Cytometry: EGFR [p Tyr1045] Antibody (PSH03-28) [NBP3-32760]

Flow Cytometry: EGFR [p Tyr1045] Antibody (PSH03-28) [NBP3-32760] - Flow cytometric analysis of A431 treated with or without 100ng/mL EGF for 60 minutes cells labeling EGFR.

Cells were fixed and permeabilized. Then stained with the primary antibody (NBP3-32760, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

Applications for EGFR [p Tyr1045] Antibody (PSH03-28)

Application
Recommended Usage

Flow Cytometry

1:1000

Immunocytochemistry/ Immunofluorescence

1:100

Western Blot

1:1000

Formulation, Preparation, and Storage

Purification

Protein A purified

Formulation

PBS (pH7.4), 0.1% BSA and 40% Glycerol

Preservative

0.05% Sodium Azide

Concentration

1 mg/ml

Shipping

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

Stability & Storage

Store at 4C short term. Aliquot and store at -20C 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

Additional EGFR Products

Product Documents for EGFR [p Tyr1045] Antibody (PSH03-28)

Product Datasheets

Certificate of Analysis

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Product Specific Notices for EGFR [p Tyr1045] Antibody (PSH03-28)

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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