• Assay Type

  Solid Phase Sandwich ELISA

• Format

  96-well strip plate

• Sample Type & Volume Required

  Cell lysates (100 µL)

• Sufficient Materials

  Kits available for two, five, or fifteen 96-well plates*

• Specificity

  Please see the

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

This DuoSet IC ELISA contains the basic components required for the development of sandwich ELISAs to measure tyrosine-phosphorylated

Product Features

• Optimized capture and detection antibody pairings and 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

Kit Content

• Capture Antibody

• Conjugated Detection Antibody

• Calibrated Immunoassay Standard or Control

Other Reagents Required

PBS: (Catalog # ), 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 # ), or equivalent

Lysis Buffer*

IC Diluent*

Blocking Buffer*

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

Stop Solution: 2 N H₂SO₄ (Catalog # )

Microplates: R&D Systems (Catalog # ), or equivalent

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

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

Preparation and Storage

• 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.

• Entrez Gene IDs:

  1956 (Human); 13649 (Mouse);

• Long Name:

  Epidermal Growth Factor Receptor

• Aliases:

  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