Human Total EGFR DuoSet IC ELISA

Name: Human Total EGFR DuoSet IC ELISA
Brand: R&D Systems
SKU: DYC1854-2
Rating: 3 (1 reviews)

Catalog # DYC1854-2 | R&D Systems, Inc. a Bio-Techne Brand

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Citations (9)

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Key Product Details

Assay Type:	Solid Phase Sandwich ELISA
Assay Range:	31.2 - 2,000 pg/mL
Sample Types:	Cell Lysates
Human Total EGFR DuoSet IC ELISA Features:	Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time Development protocols are provided to guide further assay optimization Assay can be customized to your specific needs Available in 2, 5, and 15- (96-well) plate pack sizes Economical alternative to Western blot

View Full Technical Details for Human Total EGFR DuoSet IC ELISA

Summary for Human Total EGFR DuoSet IC ELISA

This DuoSet IC ELISA contains the basic components required for the development of sandwich ELISAs to measure total EGFR in cell lysates. An immobilized capture antibody specific for EGFR binds both phosphorylated and unphosphorylated EGFR. After washing away unbound material, a biotinylated detection antibody is used to detect both phosphorylated and unphosphorylated protein, utilizing a standard Streptavidin-HRP format.

Product Specifications for Human Total EGFR DuoSet IC ELISA

Assay Format	96-well strip plate (sold separately)
Sample Volume Required	Cell lysates (100 µL)
Reactivity	Human
Specificity	Please see the product datasheet
Detection Method	Colorimetric ELISA - 450nm (TMB)
Conjugate	Biotin
Label	HRP
Measurement	Quantitative ELISA
Sufficient Materials	Kits available for two, five, or fifteen 96-well plates*

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

Kit Contents for Human Total EGFR DuoSet IC ELISA

Capture Antibody
Conjugated Detection Antibody
Calibrated Immunoassay Standard or Control
Streptavidin-HRP

Other Reagents Required

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2O4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or equivalent

Lysis Buffer*

IC Diluent*

Blocking Buffer*

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: From Costar EIA Plate (Costar Catalog # 2592) or R&D Systems (Catalog # DY990), or equivalent

Plate Sealers: ELISA Plate Sealers (Catalog # DY992), or equivalent

*For the Lysis Buffer, IC Diluent, and Blocking BUffer recommended for a specific DuoSet ELISA Development Kit, please see the product

Scientific Data Examples for Human Total EGFR DuoSet IC ELISA

	Standard Curve
	Figure 1. The Human Total-EGF R DuoSet IC ELISA is more sensitive than immunoprecipitation (IP)-Western blot analysis Lysates prepared from the human epidermoid carcinoma cell line, A431, were diluted in series and analyzed by (A) IP-Western blot and (B) R&D Systems’ Human Total-EGF R DuoSet IC ELISA (Catalog # DYC1854). IPs were performed using anti-EGF R polyclonal antibody (Catalog # AF231) Immunoblots were incubated with a biotinylated anti-EGF R polyclonal antibody (Catalog # BAF231) to detect total EGF R. Blots were incubated with Streptavidin-HRP (Catalog # DY998) followed by chemiluminescent detection. Human EGF R can be detected by the Human Total-EGF R DuoSet IC ELISA by using approximately 200 to 500 times less lysate than is needed for a conventional IP-Western blot.
	Figure 2. The Human Total-EGF R DuoSet IC ELISA measures the relative level of EGF R Lysates were prepared from A431 cells, human immortalized fibroblast cells (CCD1070SK), and human breast adenocarcinoma cells (SkBr3). ELISA analysis was done using 125 ng (A431) and 2000 ng (CCD1070SK and SkBr3) of lysate. IP-Western blot analysis (inset) was done using 5 μg (A431) and 100 μg (CCD1070SK and SkBr3) of lysate. The IP-Western blot was performed as described in Figure 1. The smaller band visible in IP-Western blot is an immature EGF R glycosylation intermediate.

Preparation and Storage

Shipping	The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage	Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: EGFR

The EGF R subfamily of receptor tyrosine kinases comprises four members: EGF R (also known as HER-1, ErbB1, or ErbB), ErbB2 (Neu, HER-2), ErbB3 (HER-3), and ErbB4 (HER-4). All family members are type I transmembrane glycoproteins with an extracellular ligand binding domain containing two cysteine-rich domains separated by a spacer region and a cytoplasmic domain containing a membrane-proximal tyrosine kinase domain followed by multiple tyrosine autophosphorylation sites. The human EGF R cDNA encodes a 1210 amino acid (aa) precursor with a 24 aa signal peptide, a 621 aa extracellular domain (ECD), a 23 aa transmembrane segment, and a 542 aa cytoplasmic domain. Soluble receptors consisting of the extracellular ligand binding domain are generated by alternate splicing in human and mouse. Within the ECD, human EGF R shares 88% aa sequence identity with mouse and rat EGF R. It shares 43% - 44% aa sequence identity with the ECD of human ErbB2, ErbB3, and ErbB4. EGF R binds a subset of the EGF family ligands, including EGF, amphiregulin, TGF-alpha, betacellulin, epiregulin, HB-EGF, and epigen. Ligand binding induces EGF R homodimerization as well as heterodimerization with ErbB2, resulting in kinase activation, heterodimerization tyrosine phosphorylation and cell signaling. EGF R can also be recruited to form heterodimers with the ligand-activated ErbB3 or ErbB4. EGF R signaling regulates multiple biological functions including cell proliferation, differentiation, motility, and apoptosis. EGF R is overexpressed in a wide variety of tumors and is the target of several anti-cancer drugs.

Item	Value
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 Total EGFR DuoSet IC ELISA (9)

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Citations are publications that use Bio-Techne products. Selected citations for Human Total EGFR DuoSet IC ELISA include:

Species: Human
Sample Types: Tissue Homogenates

N Floc'h et al. (2018-02-26), Anti-tumor activity of osimertinib, an irreversible mutant-selective EGFR tyrosine kinase inhibitor, in NSCLC harboring EGFR Exon 20 Insertions Mol. Cancer Ther., 2018-00(0).
PMID: 29483211
Species: Human
Sample Types: Cell Culture Supernates

TM Scharadin et al. (2017-06-06), Synthesis and biochemical characterization of EGF receptor in a water-soluble membrane model system PLoS ONE, 2017-012(6):e0177761.
PMID: 28586369
Species: Human
Sample Types: Cell Lysates

Adam Jedlinski (2017-01-28), Cetuximab sensitivity of head and neck squamous cell carcinoma xenografts is associated with treatment-induced reduction of EGFR, pEGFR, and pSrc J. Oral Pathol. Med, 2017-00(0).
PMID: 28036101
Species: Human
Sample Types: Urine

Bryan R et al. (2015-03-17), Protein shedding in urothelial bladder cancer: prognostic implications of soluble urinary EGFR and EpCAM. Br J Cancer, 2015-0112(6):1052-8.
PMID: 25719831
Species: Human
Sample Types: Cell Lysates

Kim S et al. (2011-09-07), Epidermal growth factor receptor reactivation induced by E-prostanoid-3 receptor- and tumor necrosis factor-alpha-converting enzyme-dependent feedback exaggerates interleukin-8 production in airway cancer (NCI-H292) cells. Exp. Cell Res., 2011-0317(18):2650-60.
PMID: 21925169
Species: Human
Sample Types: Tissue Homogenates

Yamashita-Kashima Y et al. (2011-06-23), Pertuzumab in combination with trastuzumab shows significantly enhanced antitumor activity in HER2-positive human gastric cancer xenograft models. Clin. Cancer Res., 2011-017(15):5060-70.
PMID: 21700765
Species: Human
Sample Types: Cell Lysates

O'Brien NA et al. (2010-05-25), Activated phosphoinositide 3-kinase/AKT signaling confers resistance to trastuzumab but not lapatinib. Mol. Cancer Ther., 2010-09(6):1489-502.
PMID: 20501798
Species: Human
Sample Types: Cell Lysates

Hickinson DM et al. (2009-06-01), Identification of biomarkers in human head and neck tumor cell lines that predict for in vitro sensitivity to gefitinib. Clin Transl Sci, 2009-02(3):183-92.
PMID: 20443891
Species: Human
Sample Types: Cell Lysates

Shankaran H et al. (2008-04-18), Quantifying the effects of co-expressing EGFR and HER2 on HER activation and trafficking. Biochem. Biophys. Res. Commun., 2008-0371(2):220-4.
PMID: 18424261

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Human Total EGFR DuoSet IC ELISA

Name: Balaji Mahender

Sample Tested: Cell culture supernatant

Verified Customer | Posted 01/18/2018

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Product Documents for Human Total EGFR DuoSet IC ELISA

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Human Total EGFR DuoSet IC ELISA

Catalog # DYC1854-2 | R&D Systems, Inc. a Bio-Techne Brand

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Summary for Human Total EGFR DuoSet IC ELISA

Product Specifications for Human Total EGFR DuoSet IC ELISA

Kit Contents for Human Total EGFR DuoSet IC ELISA

Other Reagents Required

Scientific Data Examples for Human Total EGFR DuoSet IC ELISA

Preparation and Storage

Background: EGFR

Citations for Human Total EGFR DuoSet IC ELISA (9)

Customer Reviews for Human Total EGFR DuoSet IC ELISA (1)

Human Total EGFR DuoSet IC ELISA

Product Documents for Human Total EGFR DuoSet IC ELISA

Product Datasheets

COA

Certificate of Analysis

SDS

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