Human EGFR Antibody

Catalog # Availability Size / Price Qty
AF231
AF231-SP
Detection  of Human EGFR by Western Blot.
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Product Details
Citations (8)
FAQs
Supplemental Products
Reviews (1)

Human EGFR Antibody Summary

Species Reactivity
Human
Specificity
Detects human EGFR in ELISAs and Western blots. In sandwich ELISAs, approximately 3% cross-reactivity with recombinant mouse EGFR is observed and less than 0.1% cross-reactivity with recombinant human (rh) ErbB2 and rhErbB3 is observed.
Source
Polyclonal Goat IgG
Purification
Antigen Affinity-purified
Immunogen
Mouse myeloma cell line NS0-derived recombinant human EGFR
Leu25-Ser645
Accession # CAA25240
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
1 µg/mL
See below
Flow Cytometry
0.25 µg/106 cells
See below
Immunohistochemistry
1-15 µg/mL
See below
Immunoprecipitation
1 µg/mL
A431 human epithelial carcinoma cell line, see our available Western blot detection antibodies
CyTOF-ready
Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
 
Immunocytochemistry
1-15 µg/mL
See below

Human EGFR Sandwich Immunoassay

Recommended Concentration
Reagent
ELISA Capture (Matched Antibody Pair)
0.2-0.8 µg/mL 

Use in combination with:

Detection Reagent: Human EGFR Biotinylated Antibody (Catalog # BAF231)

Standard: Recombinant Human EGFR Protein (Catalog # 1095-ER)

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Data Examples

Western Blot Detection  of Human EGFR by Western Blot. View Larger

Detection of Human EGFR by Western Blot. Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line and MDA‑MB‑231 human breast cancer cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human EGFR Antigen Affinity-purified Polyclonal Antibody (Catalog # AF231) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for EGFR at approximately 175 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Flow Cytometry Detection  of EGFR in A431 Human Cell Line by Flow Cytometry. View Larger

Detection of EGFR in A431 Human Cell Line by Flow Cytometry. A431 human epithelial carcinoma cell line was stained with Goat Anti-Human EGFR Antigen Affinity-purified Polyclonal Antibody (Catalog # AF231, filled histogram) or isotype control antibody (Catalog # AB-108-C, open histogram), followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0107). View our protocol for Staining Membrane-associated Proteins.

Immunocytochemistry EGFR  in A431 Human Cell Line. View Larger

EGFR in A431 Human Cell Line. EGFR was detected in immersion fixed A431 human epithelial carcinoma cell line using Goat Anti-Human EGFR Antigen Affinity-purified Polyclonal Antibody (Catalog # AF231) at 1 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to plasma membrane. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Immunohistochemistry EGFR  in Human Skin. View Larger

EGFR in Human Skin. EGFR was detected in immersion fixed frozen sections of human skin using Goat Anti-Human EGFR Antigen Affinity-purified Polyclonal Antibody (Catalog # AF231) at 1 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC004). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to plasma membrane. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: EGFR

The epidermal growth factor receptor (EGFR) subfamily of receptor tyrosine kinases comprises four members: EGFR (also known as HER1, ErbB1 or ErbB), ErbB2 (Neu, HER2), ErbB3 (HER3), and ErbB4 (HER4). All family members are type I transmembrane glycoproteins that have an extracellular domain which contains two cysteine-rich domains separated by a spacer region that is involved in ligand binding, and a cytoplasmic domain which has a membrane-proximal tyrosine kinase domain and a C-terminal tail with multiple tyrosine autophosphorylation sites. The human EGFR gene encodes a 1210 amino acid (aa) residue precursor with a 24 aa putative signal peptide, a 621 aa extracellular domain, a 23 aa transmembrane domain, and a 542 aa cytoplasmic domain. EGFR has been shown to bind a subset of the EGF family ligands, including EGF, amphiregulin, TGF-alpha, betacellulin, epiregulin, heparin-binding EGF and neuregulin-2 alpha  in the absence of a co-receptor. Ligand binding induces EGFR homodimerization as well as heterodimerization with ErbB2, resulting in kinase activation, tyrosine phosphorylation and cell signaling. EGFR can also be recruited to form heterodimers with the ligand-activated ErbB3 or ErbB4. EGFR signaling has been shown to regulate multiple biological functions including cell proliferation, differentiation, motility and apoptosis. In addition, EGFR signaling has also been shown to play a role in carcinogenesis (1 - 3).

References
  1. Daly, R.J. (1999) Growth Factors, 16:255.
  2. Schlessinger, J. (2000) Cell. 103:211.
  3. Maihle, N.J. et al. (2002) Cancer Treat. Res. 107:247.
Long Name
Epidermal Growth Factor Receptor
Entrez Gene IDs
1956 (Human); 13649 (Mouse)
Alternate Names
avian erythroblastic leukemia viral (v-erb-b) oncogene homolog; cell growth inhibiting protein 40; cell proliferation-inducing protein 61; EC 2.7.10; EC 2.7.10.1; EGF R; EGFR; epidermal growth factor receptor (avian erythroblastic leukemia viral (v-erb-b)oncogene homolog); epidermal growth factor receptor; ErbB; ErbB1; ERBB1PIG61; HER1; HER-1; mENA; Proto-oncogene c-ErbB-1; Receptor tyrosine-protein kinase erbB-1

Product Datasheets

Citations for Human EGFR Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

8 Citations: Showing 1 - 8
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  1. Improved efficiency of in situ protein analysis by proximity ligation using UnFold probes
    Authors: A Klaesson, K Grannas, T Ebai, J Heldin, B Koos, M Leino, D Raykova, J Oelrich, L Arngården, O Söderberg, U Landegren
    Sci Rep, 2018;8(1):5400.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC-P
  2. Identification of a novel anoikis signalling pathway using the fungal virulence factor gliotoxin
    Authors: F Haun, S Neumann, L Peintner, K Wieland, J Habicht, C Schwan, K Østevold, MM Koczorowsk, M Biniossek, M Kist, H Busch, M Boerries, RJ Davis, U Maurer, O Schilling, K Aktories, C Borner
    Nat Commun, 2018;9(1):3524.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  3. Soluble fms-like tyrosine kinase 1 promotes angiotensin II sensitivity in preeclampsia
    Authors: Suzanne D Burke
    J Clin Invest, 2016;0(0):.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC-P
  4. Ability of the Met kinase inhibitor crizotinib and new generation EGFR inhibitors to overcome resistance to EGFR inhibitors.
    Authors: Nanjo, Shigeki, Yamada, Tadaaki, Nishihara, Hiroshi, Takeuchi, Shinji, Sano, Takako, Nakagawa, Takayuki, Ishikawa, Daisuke, Zhao, Lu, Ebi, Hiromich, Yasumoto, Kazuo, Matsumoto, Kunio, Yano, Seiji
    PLoS ONE, 2013;8(12):e84700.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  5. Human epidermal growth factor receptor (HER-1:HER-3) Fc-mediated heterodimer has broad antiproliferative activity in vitro and in human tumor xenografts.
    Authors: Sarup J, Jin P, Turin L, Bai X, Beryt M, Brdlik C, Higaki JN, Jorgensen B, Lau FW, Lindley P, Liu J, Ni I, Rozzelle J, Kumari R, Watson SA, Zhang J, Shepard HM
    Mol. Cancer Ther., 2008;7(10):3223-36.
    Species: Human
    Sample Types: Cell Lysates
    Applications: ELISA Development
  6. Development and validation of sandwich ELISA microarrays with minimal assay interference.
    Authors: Gonzalez RM, Seurynck-Servoss SL, Crowley SA
    J. Proteome Res., 2008;7(6):2406-14.
    Species: Human
    Sample Types: Serum
    Applications: ELISA Microarray Development
  7. FGFR2-amplified gastric cancer cell lines require FGFR2 and Erbb3 signaling for growth and survival.
    Authors: Kunii K, Davis L, Gorenstein J, Hatch H, Yashiro M, Di Bacco A, Elbi C, Lutterbach B
    Cancer Res., 2008;68(7):2340-8.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Immunoprecipitation
  8. Expression of growth factors and growth factor receptor in non-healing and healing ischaemic ulceration.
    Authors: Murphy MO, Ghosh J, Fulford P, Khwaja N, Halka AT, Carter A, Turner NJ, Walker MG
    Eur J Vasc Endovasc Surg, 2006;31(5):516-22.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC

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Human EGFR Antibody
By Anonymous on 07/31/2018
Application: WB Sample Tested: HT-29 human colon adenocarcinoma cell line Species: Human