Human EGFR Antibody

Name: Human EGFR Antibody
Brand: R&D Systems
SKU: AF231
Rating: 5 (1 reviews)

R&D Systems, Inc. a Bio-Techne Brand

Catalog # AF231

Citations (15)

(1)

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Key Product Details

Species Reactivity:	Human
Immunogen:	Mouse myeloma cell line NS0-derived recombinant human EGFR Leu25-Ser645 Accession # CAA25240
Applications:	CyTOF-ready, ELISA Capture, Flow Cytometry, Immunocytochemistry/Immunofluorescence, Immunohistochemistry, Immunoprecipitation, Simple Western, Western Blot
Label:	Unconjugated
Source:	Polyclonal Goat IgG
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.

View Full Technical Details for Human EGFR Antibody

Applications for Human EGFR Antibody

	Recommended Concentration	Sample
Western Blot	1 µg/mL	See below
Simple Western	12.5 µg/mL	A431 human epithelial carcinoma cell line
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.

Scientific Data Examples for Human EGFR Antibody

	Western Blot 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. 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. 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. 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.
	Simple Western Detection of Human EGFR by Simple Western^TM. Simple Western lane view shows lysates of A431 human epithelial carcinoma cell line, loaded at 4.2 mg/mL. A specific band was detected for EGFR at approximately 229 kDa (as indicated) using 12.5 µg/mL of Goat Anti-Human EGFR Antigen Affinity-purified Polyclonal Antibody (Catalog # AF231). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Formulation, Preparation and Storage

Purification	Antigen Affinity-purified
Reconstitution	Reconstitute at 0.2 mg/mL in sterile PBS. Reconstitution Buffer Available: Size / Price Qty Loading... RB01:
Formulation	Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
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.

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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).

Item	Value
References	Daly, R.J. (1999) Growth Factors, 16:255. Schlessinger, J. (2000) Cell. 103:211. Maihle, N.J. et al. (2002) Cancer Treat. Res. 107:247.
Long Name	Epidermal Growth Factor Receptor
Entrez Gene IDs	1956 (Human); 13649 (Mouse); 24329 (Rat); 102138724 (Cynomolgus Monkey)
Alternate Names	EC 2.7.10, EC 2.7.10.1, EGF R, EGFR, ERBB1PIG61, ErbB, ErbB1, HER-1, HER1, Proto-oncogene c-ErbB-1, Receptor tyrosine-protein kinase erbB-1, avian erythroblastic leukemia viral (v-erb-b) oncogene homolog, cell growth inhibiting protein 40, cell proliferation-inducing protein 61, epidermal growth factor receptor, epidermal growth factor receptor (avian erythroblastic leukemia viral (v-erb-b)oncogene homolog), mENA

Citations for Human EGFR Antibody (15)

Showing 1 - 10 of 15 citations Showing All

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Citations are publications that use Bio-Techne products. Selected citations for Human EGFR Antibody include:

Species: Human
Sample Types: Cell Lysates
Applications: Western Blot

C Hu et al. (2022), Glioblastoma mutations alter EGFR dimer structure to prevent ligand bias Nature, 602(7897):518-522.
PMID: 35140400
Species: Human
Sample Types: Cell Lysates
Applications: Immunoprecipitation

DM Beauvais et al. (2022), Plasma membrane proteoglycans syndecan-2 and syndecan-4 engage with EGFR and RON kinase to sustain carcinoma cell cycle progression The Journal of Biological Chemistry, 0(0):102029.
PMID: 35569509
Species: Mouse
Sample Types: Whole Tissue
Applications: IHC

S Sugita et al. (2021), Involvement of cancer-derived EMT cells in the accumulation of 18F-fluorodeoxyglucose in the hypoxic cancer microenvironment Scientific Reports, 11(1):9668.
PMID: 33994540
Species: Human
Sample Types: Cell Lysates
Applications: Co-Immunoprecipitation

X Liu et al. (2021), EGFR inhibition blocks cancer stem cell clustering and lung metastasis of triple negative breast cancer Theranostics, 11(13):6632-6643.
PMID: 33995681
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot

R Chakrabort et al. (2021), Bacterial Antigens Reduced the Inhibition Effect of Capsaicin on Cal 27 Oral Cancer Cell Proliferation International Journal of Molecular Sciences, 22(16).
PMID: 34445392
Species: Human
Sample Types: Whole Cell
Applications: IF

T Egnuni et al. (2021), Evaluation of the Targeting and Therapeutic Efficiency of Anti-EGFR Functionalised Nanoparticles in Head and Neck Cancer Cells for Use in NIR-II Optical Window Pharmaceutics, 13(10).
PMID: 34683944
Species: Human
Sample Types: Cell Lysates
Applications: ELISA Detection

N Yogo et al. (2020), Development of an immuno-wall device for the rapid and sensitive detection of EGFR mutations in tumor tissues resected from lung cancer patients PLoS ONE, 15(11):e0241422.
PMID: 33196648
Species: Human
Sample Types: Whole Tissue
Applications: IHC-P

A Klaesson et al. (2018), Improved efficiency of in situ protein analysis by proximity ligation using UnFold probes Sci Rep, 8(1):5400.
PMID: 29599435
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot

F Haun et al. (2018), Identification of a novel anoikis signalling pathway using the fungal virulence factor gliotoxin Nat Commun, 9(1):3524.
PMID: 30166526
Species: Human
Sample Types: Whole Tissue
Applications: IHC-P

Suzanne D Burke (2016), Soluble fms-like tyrosine kinase 1 promotes angiotensin II sensitivity in preeclampsia J Clin Invest, 0(0).
PMID: 27270170
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot

Nanjo et al. (2013), Ability of the Met kinase inhibitor crizotinib and new generation EGFR inhibitors to overcome resistance to EGFR inhibitors. PLoS ONE, 8(12):e84700.
PMID: 24386407
Species: Human
Sample Types: Cell Lysates
Applications: ELISA Development

Sarup J et al. (2008), Human epidermal growth factor receptor (HER-1:HER-3) Fc-mediated heterodimer has broad antiproliferative activity in vitro and in human tumor xenografts. Mol. Cancer Ther., 7(10):3223-36.
PMID: 18852126
Species: Human
Sample Types: Serum
Applications: ELISA Microarray Development

Gonzalez RM et al. (2008), Development and validation of sandwich ELISA microarrays with minimal assay interference. J. Proteome Res., 7(6):2406-14.
PMID: 18422355
Species: Human
Sample Types: Cell Lysates
Applications: Immunoprecipitation

Kunii K et al. (2008), FGFR2-amplified gastric cancer cell lines require FGFR2 and Erbb3 signaling for growth and survival. Cancer Res., 68(7):2340-8.
PMID: 18381441
Species: Human
Sample Types: Whole Tissue
Applications: IHC

Murphy MO et al. (2006), Expression of growth factors and growth factor receptor in non-healing and healing ischaemic ulceration. Eur J Vasc Endovasc Surg, 31(5):516-22.
PMID: 16427789

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Customer Reviews for Human EGFR Antibody (1)

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Human EGFR Antibody

Name: Anonymous

Application: WB

Sample Tested: HT-29 human colon adenocarcinoma cell line

Species: Human

Verified Customer | Posted 07/31/2018

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Product Documents for Human EGFR Antibody

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Human EGFR Antibody

R&D Systems, Inc. a Bio-Techne Brand

Key Product Details

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Applications for Human EGFR Antibody

Human EGFR Sandwich Immunoassay

Scientific Data Examples for Human EGFR Antibody

Western Blot

Flow Cytometry

Immunocytochemistry

Immunohistochemistry

Simple Western

Formulation, Preparation and Storage

Reconstitution Calculator

Background: EGFR

Citations for Human EGFR Antibody (15)

Customer Reviews for Human EGFR Antibody (1)

Customer Images

Human EGFR Antibody

Product Documents for Human EGFR Antibody

Product Datasheets

COA

Certificate of Analysis

SDS

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