Human Phospho-EGFR DuoSet IC ELISA, 2 Plate

The human epidermoid carcinoma cell line, A431, was treated with 25 ng/mL recombinant human EGF (R&D Systems Catalog #236-EG) for 5 minutes to induce tyrosine phosphorylation of EGF R. Lysates were serially diluted and analyzed by (A) IP-Western blot and (B) this DuoSet IC ELISA. IPs were done using an anti-EGF R monoclonal antibody and anti-mouse IgG agarose. Immunoblots were incubated with an HRP-conjugated anti-phospho-tyrosine monoclonal antibody (R&D Systems, Catalog #HAM1676) to detect phosphor-EGF R. Bands were visualized by chemiluminescent detection. Human Phospho-EGF R can be detected in this DuoSet IC ELISA by using approximately 2-4 times less lysate than is needed for a conventional IP-Western blot.

A431 cells were untreated or treated with 25 ng/mL recombinant human EGF for 5 minutes. ELISA and IP-Western blot (inset) analyses were done using 25 and 50 μg of lysate, respectively. IP-Western blots for phospho-EGF R (p-EGF R) were done as described in Figure 1. Blots were stripped and total EGF R was detected using a biotinylated anti-EGF R polyclonal antibody (R&D Systems, Catalog #AF231).

A431 cells were incubated with no additions or treated with 25 ng/mL recombinant human EGF for 5 minutes, after either being treated with or without 1 nM, 10 nM, or 100 nM PD168393. ELISA and IP-Western blot (inset) analyses were done using 25 and 50 μg of lysate, respectively. IP-Western blots for phospho-EGF R (p-EGF R) were done as described in Figure 1. Total EGF R blots were done as described in Figure 2. The DuoSet IC ELISA results correlate well with the relative amount of phosphorylated EGF R detected by IP-Western blot.

A431 cells were treated with 25 ng/mL recombinant human EGF for 5 minutes. The indicated amounts of recombinant extracellular domains of human EGF R (R&D Systems, Catalog #1095-ER), human ErbB2/Fc Chimera (R&D Systems, Catalog #1129-ER), human ErbB3/Fc Chimera (R&D Systems, Catalog #348-RB), or human ErbB4/Fc Chimera (R&D Systems, Catalog #1131-ER) were added to 25 μg of lysate and analyzed using this DuoSet IC ELISA. Competition was observed only with recombinant human EGF R.

• Reactivity: Hu
• Applications: ELISA
• Conjugate: Biotin

Human Phospho-EGFR DuoSet IC ELISA, 2 Plate Summary

• Source: N/A
• Assay Type: Solid Phase Sandwich ELISA
• Inter-Assay: See PDF Datasheet for details
• Intra-Assay: See PDF Datasheet for details
• Spike Recovery: See PDF Datasheet for details
• Sample Volume: See PDF Datasheet for details
• Gene: EGFR

Applications/Dilutions

• Dilutions:
  • ELISA
• Application Notes: No significant interference observed with available related molecules.

Packaging, Storage & Formulations

• Storage: Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Human Phospho-EGFR DuoSet IC ELISA, 2 Plate

• 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

Background

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.

Publications for EGFR (DYC1095B-2)(14)

Publications using DYC1095B-2

• N Floc'h, MJ Martin, JW Riess, JP Orme, AD Staniszews, L Menard, ME Cuomo, DJ O'Neill, RA Ward, MRV Finlay, D McKerreche, M Cheng, DP Vang, RA Tsai, JG Keck, DR Gandara, PC Mack, DA Cross Anti-tumor activity of osimertinib, an irreversible mutant-selective EGFR tyrosine kinase inhibitor, in NSCLC harboring EGFR Exon 20 Insertions Mol. Cancer Ther., 2018-02-26;0(0):. 2018-02-26 [PMID: 29483211] (Human)
• M Su, B Qin, F Liu, Y Chen, R Zhang Andrographolide enhanced 5-fluorouracil-induced antitumor effect in colorectal cancer via inhibition of c-MET pathway Drug Des Devel Ther, 2017-11-23;11(0):3333-3341. 2017-11-23 [PMID: 29200829] (Human)
• Adam Jedlinski 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, 2017-01-28;0(0):. 2017-01-28 [PMID: 28036101] (Human)
• Stand-Sit Microchip for High-Throughput, Multiplexed Analysis of Single Cancer Cells Sci Rep, 2016-09-01;6(0):32505. 2016-09-01 [PMID: 27581736] (ChIP)
• Ekerljung L, Lennartsson J, Gedda L The HER2-binding affibody molecule (Z(HER2ratio342))(2) increases radiosensitivity in SKBR-3 cells. PLoS ONE, 2012-11-14;7(11):e49579. 2012-11-14 [PMID: 23166716] (Human)
• Brave SR, Odedra R, James NH Vandetanib inhibits both VEGFR-2 and EGFR signalling at clinically relevant drug levels in preclinical models of human cancer. Int. J. Oncol., 2011-04-29;39(1):271-8. 2011-04-29 [PMID: 21537841] (Human)
• Giorgio C, Hassan Mohamed I, Flammini L, Barocelli E, Incerti M, Lodola A, Tognolini M Lithocholic acid is an Eph-ephrin ligand interfering with Eph-kinase activation. PLoS ONE, 2011-03-30;6(3):e18128. 2011-03-30 [PMID: 21479221] (Human)
• Kim S, Lewis C, Nadel JA CCL20/CCR6 feedback exaggerates epidermal growth factor receptor-dependent MUC5AC mucin production in human airway epithelial (NCI-H292) cells. J. Immunol., 2011-02-07;186(6):3392-400. 2011-02-07 [PMID: 21300824] (Human)
• O'Brien NA, Browne BC, Chow L, Wang Y, Ginther C, Arboleda J, Duffy MJ, Crown J, O'Donovan N, Slamon DJ Activated phosphoinositide 3-kinase/AKT signaling confers resistance to trastuzumab but not lapatinib. Mol. Cancer Ther., 2010-05-25;9(6):1489-502. 2010-05-25 [PMID: 20501798] (Human)
• Hickinson DM, Klinowska T, Speake G AZD8931, an equipotent, reversible inhibitor of signaling by epidermal growth factor receptor, ERBB2 (HER2), and ERBB3: a unique agent for simultaneous ERBB receptor blockade in cancer. Clin. Cancer Res., 2010-02-09;16(4):1159-69. 2010-02-09 [PMID: 20145185] (Human)
• Kim S, Nadel JA Fibrinogen binding to ICAM-1 promotes EGFR-dependent mucin production in human airway epithelial cells. Am. J. Physiol. Lung Cell Mol. Physiol., 2009-05-08;297(1):L174-83. 2009-05-08 [PMID: 19429776] (Human)
• Shankaran H, Zhang Y, Opresko L, Resat H Quantifying the effects of co-expressing EGFR and HER2 on HER activation and trafficking. Biochem. Biophys. Res. Commun., 2008-04-18;371(2):220-4. 2008-04-18 [PMID: 18424261] (Human)
• Liu M, Yang SC, Sharma S, Luo J, Cui X, Peebles KA, Huang M, Sato M, Ramirez RD, Shay JW, Minna JD, Dubinett SM EGFR signaling is required for TGF-beta 1 mediated COX-2 induction in human bronchial epithelial cells. Am. J. Respir. Cell Mol. Biol., 2007-06-28;37(5):578-88. 2007-06-28 [PMID: 17600311] (Human)
• Sakai K, Yokote H, Murakami-Murofushi K, Tamura T, Saijo N, Nishio K In-frame deletion in the EGF receptor alters kinase inhibition by gefitinib. Biochem. J., 2006-08-01;397(3):537-43. 2006-08-01 [PMID: 16623663] (Human)

FAQs for EGFR (DYC1095B-2). (Showing 1 - 1 of 1 FAQs).

1. wondering what the difference is between your quantikine and duo set elisas?
   • Usually the duosets do not have the entire kit such as plates and buffers, whereas the other kits are complete.

Bioinformatics

• Gene Symbol: EGFR