• 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: Tie-2

Tie-1/Tie and Tie-2/Tek are receptor tyrosine kinases with unique structural characteristics including two immunoglobulin-like domains flanking three epidermal growth factor (EGF)-like domains, followed by three fibronectin type III-like repeats in the extracellular region, and a split tyrosine kinase domain in the cytoplasmic region. Tie-2 is involved in vascular stabilization and remodeling. Although less well understood, Tie-1 may also act as an ANG receptor, possibly in complex with Tie-2.

Human Tie-2 cDNA encodes a 1124 amino acid (aa) residue precursor protein with an 18 residue putative signal peptide, a 727 residue extracellular domain and a 354 residue cytoplasmic domain. Tie-2 is a receptor for the angiopoietin (ANG) family: ANG-1, ANG-2, and ANG-3 (mouse)/-4 (human). Ang-2 has been reported to act as an antagonist for Ang-1. Mice engineered to overexpress Ang-2 or to lack Ang-1 or Tie-2 display similar angiogenesis defects.

• Entrez Gene IDs:

  7010 (Human); 21687 (Mouse); 30747 (Zebrafish);

• Long Name:

  Tyrosine Kinase with Immunoglobulin and Epidermal Growth Factor Homology Domains 2

• Aliases:

  angiopoietin-1 receptor; CD202b antigen; CD202b; EC 2.7.10; EC 2.7.10.1; hTIE2; p140 TEK; soluble TIE2 variant 1; soluble TIE2 variant 2; TEK tyrosine kinase, endothelial; TEK; Tie2; Tie-2; TIE2CD202b; Tunica interna endothelial cell kinase; Tyrosine-protein kinase receptor TEK; Tyrosine-protein kinase receptor TIE-2; venous malformations, multiple cutaneous and mucosal; VMCM; VMCM1