EGFR Antibody (PD00-75)
Novus Biologicals, part of Bio-Techne | Catalog # NBP3-32255
Recombinant Monoclonal Antibody
Key Product Details
Species Reactivity
Human
Applications
Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence
Label
Unconjugated
Antibody Source
Recombinant Monoclonal Rabbit IgG Clone # PD00-75 expressed in HEK293
Concentration
1 mg/ml
Product Specifications
Immunogen
Synthetic peptide corresponding to C-terminus of human EGFR protein. (Uniprot: P00533)
Localization
Cell membrane, Endoplasmic reticulum membrane, Golgi apparatus membrane, Nucleus membrane, Endosome, Nucleus, Secreted.
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.
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 Antibody (PD00-75)
Immunohistochemistry: EGFR Antibody (PD00-75) [NBP3-32255]
Immunohistochemistry: EGFR Antibody (PD00-75) [NBP3-32255] - Immunohistochemical analysis of paraffin-embedded human skin tissue with Rabbit anti-EGFR antibody (NBP3-32255) at 1/800 dilution.The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (NBP3-32255) at 1/800 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunocytochemistry/ Immunofluorescence: EGFR Antibody (PD00-75) [NBP3-32255]
Immunocytochemistry/ Immunofluorescence: EGFR Antibody (PD00-75) [NBP3-32255] - Immunocytochemistry analysis of A431 cells labeling EGFR with Rabbit anti-EGFR antibody (NBP3-32255) at 1/500 dilution.Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS 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 at 1/500 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.
Flow Cytometry: EGFR Antibody (PD00-75) [NBP3-32255]
Flow Cytometry: EGFR Antibody (PD00-75) [NBP3-32255] - Flow cytometric analysis of A431 cells labeling EGFR.Cells were fixed and permeabilized. Then stained with the primary antibody (NBP3-32255, red) at 1/1,000 dilution, compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with an iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabeled sample was used as a control (cells without incubation with primary antibody; black).
Applications for EGFR Antibody (PD00-75)
Application
Recommended Usage
Flow Cytometry
1:1000
Immunocytochemistry/ Immunofluorescence
1:500
Immunohistochemistry-Paraffin
1:800
Western Blot
1:2000
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
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 (PD00-75)
Product Specific Notices for EGFR Antibody (PD00-75)
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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