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EGFR Antibody (528) - Chimeric - Azide and BSA Free

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-52651

Recombinant Monoclonal Antibody.
Novus Biologicals, part of Bio-Techne
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NBP2-52651-0.025mg
NBP2-52651-0.2mg

Key Product Details

Species Reactivity

Human

Applications

CyTOF-ready, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Paraffin, Immunoprecipitation, Neutralization, Western Blot

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Kappa Clone # 528

Format

Azide and BSA Free

Concentration

1 mg/ml

Product Specifications

Immunogen

Partially purified EGFR from A431 cells.

Specificity

Recognizes the extracellular domain of EGFR.

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG Kappa

Scientific Data Images for EGFR Antibody (528) - Chimeric - Azide and BSA Free

Immunocytochemistry/Immunofluorescence: EGFR Antibody (528) - Chimeric [NBP2-52651] -

Immunocytochemistry/Immunofluorescence: EGFR Antibody (528) - Chimeric [NBP2-52651] - Immunofluoresence staining of fixed HeLa cells with anti-EGFR antibody 528. Immunofluorescence analysis of paraformaldehyde fixed HeLa cells, permeabilized with 0.15% Triton and stained with the chimeric rabbit IgG version (NBP2-52651) at 10ug/ml for 1h followed by Alexa Fluor 488 secondary antibody (1ug/ml), showing membrane staining. The nuclear stain is DAPI (blue). Panels show from left-right, top-bottom NBP2-52651, DAPI, merged channels and a negative control. The negative control was stained with unimmunized rabbit IgG followed by Alexa Fluor 488 secondary antibody.
Flow Cytometry: EGFR Antibody (528) - Chimeric - Azide and BSA Free [NBP2-52651]

Flow Cytometry: EGFR Antibody (528) - Chimeric - Azide and BSA Free [NBP2-52651]

Flow Cytometry: EGFR Antibody (528) - Chimeric [NBP2-52651] - Flow-cytometry using the anti-EGFR antibody 528. HeLa (A) and A431 (B) cells were stained with unimmunized rabbit IgG antibody (black line) or the rabbit-chimeric version of 528 (NBP2-52651, blue line) at a concentration of 10 ug/ml for 30 mins at RT. After washing, bound antibody was detected using an anti-rabbit IgG JK (FITC-conjugate) antibody at 2 ug/ml and cells analysed on a FACSCanto flow-cytometer.

Applications for EGFR Antibody (528) - Chimeric - Azide and BSA Free

Application
Recommended Usage

Flow Cytometry

1:10 - 1:1000

Immunohistochemistry-Paraffin

1:10 - 1:500

Immunoprecipitation

1:10 - 1:500

Western Blot

1:100 - 1:2000
Application Notes
This antibody is Cytof ready. This chimeric rabbit antibody was made using the variable domain sequences of the original murine IgG2a format, for improved compatibility with existing reagents, assays and techniques.

Formulation, Preparation, and Storage

Purification

Protein A purified

Formulation

PBS

Format

Azide and BSA Free

Preservative

0.02% Proclin 300

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 for up to 3 months. For longer storage, aliquot and store at -20C.

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 Antibody (528) - Chimeric - Azide and BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for EGFR Antibody (528) - Chimeric - Azide and 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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