Human Phospho-Tie-2 DuoSet IC ELISA, 2 Plate

Human Tie-2 transfected Baf/3 cells (Baf/3-hTie-2) were treated with 600 ng/mL recombinant human Ang-1 for seven minutes to induce tyrosine phosphorylation of Tie-2. Serial dilutions of lysates were analyzed by (A) IP-Western blot and (B) this ELISA. IPs were done using an anti-Tie-2 monoclonal antibody and goat anti-mouse agarose. Immunoblots were incubated with a biotinylated anti-phosphotyrosine monoclonal antibody (Catalog # BAM1676) to detect phospho-Tie-2. Bands were visualized with Streptavidin-HRP (Cat # DY998) followed by chemiluminescent detection using WesternGloTM Chemiluminescent Detection Substrate (Catalog # AR004).

Baf/3-hTie-2 cells were untreated or treated with 600 ng/mL recombinant human Ang-1 for seven minutes. ELISA and IP-Western blot (inset) analyses were done using 50 μg and 200 μg of lysate, respectively. IP-Western blots for phospho-Tie-2 (p-Tie-2) were done as described in Figure 1. Blots were stripped and total Tie-2 was detected using a biotinylated anti-Tie-2 polyclonal antibody (Catalog # BAF313). Human Phospho-Tie-2 can be detected in this ELISA by using approximately 4 times less lysate than is needed for a conventional IP-Western blot.

Baf/3-hTie-2 cells were treated with 600 ng/mL recombinant human Ang-1 for seven minutes. The indicated amounts of recombinant extracellular domains of human Tie-2(Catalog #313-TI), human Tie-1 (Catalog #619-TI), human VEGF R2 (Catalog #357-KD) or human VEGF R3 (Catalog #321-FL) were added to 50 μg lysate and analyzed using this ELISA. Competition was observed only with recombinant Tie-2.

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

Human Phospho-Tie-2 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: TEK

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-Tie-2 DuoSet IC ELISA, 2 Plate

• 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

Background

Tie-2, also known as Tek, is a transmembrane receptor tyrosine kinase (RTK) that functions as a receptor for Angiopoietin family proteins. It is expressed by embryonic and adult endothelial cells, hematopoietic stem cells, and a circulating population of proangiogenic monocytes. Tie-2 signaling is activated by Angiopoietins-1 and -4, while it can be either activated or inhibited by Angiopoietins-2 and -3. Tie-2 plays an important role in maintaining vascular integrity by mediating endothelial cell-smooth muscle cell communication and inhibiting endothelial cell apoptosis. It is also required for embryonic development of the endocardium. In addition, Tie-2 signaling mediates the quiescence of bone marrow stem cells in response to osteoblast-produced Angiopoietin-1. This quiescence is critical for maintaining an ongoing hematopoietic capability in the bone marrow.

FAQs for Tie-2 (DYC2720-2). (Showing 1 - 1 of 1 FAQ).

1. What is the difference between the Quantikine and Duoset ELISA kits?
   • Our Quantikine kits are fully optimized and validated for the sample types listed on the product specific webpage and datasheet, as this can vary between kits. Each kit is supplied ready to use with one pre-coated 96-well plate, a detection antibody directly conjugated to HRP, and all other necessary reagents. These kits are ideal for researchers who want the convenience of a ready to use and optimized ELISA product. Some of these kits are also available prepackaged in larger 6 and 50 plate sizes.Our DuoSet Kits, in contrast, are ELISA development kits containing the capture and detection antibody, the mass-value calibrated standard, and streptavidin-HRP to prepare approximately 5 or 15 plates. Ancillary reagents will need to be used/purchased, and for most kits, we will recommend one of our Ancillary Reagent Kits, which contain the reagents we use ourselves in-house. DuoSet kits are validated only for cell culture supernatant samples and therefore require further development and validation for accurate measurement in more complex samples such as serum and plasma. Our DuoSet Kits offer an economical, flexible alternative for the experienced ELISA user.

Bioinformatics

• Gene Symbol: TEK