Human TIE2 ELISA Kit

Catalog No.: BEK1207

Size: 96T

Range: 156 pg/ml-10,000 pg/ml

Sensitivity < 5 pg/ml

Storage and Expiration: Store at 2-8℃ for 6 months, or at -20℃ for 12 months. 

Application: For quantitative detection of TIE2 in human serum, plasma, body fluids, tissue lysates or cell culture supernatants.

Introduction

Tyrosine kinase with Ig and EGF homology domain 2 (Tie-2), also called TEK tyrosine kinase, endothelial (TEK). The TEK receptor tyrosine kinase is expressed almost exclusively in endothelial cells in mice, rats, and humans. This receptor possesses a unique extracellular domain containing 2 immunoglobulin-like loops separated by 3 epidermal growth factor-like repeats that are connected to 3 fibronectin type III-like repeats. The TIE2 expression is increased in goiter in both humans and rats, consistent with a role in goitrogenesis. And the activating mutation in TIE2 caused inherited venous malformations in the 2 families and that the TIE2 signaling pathway is critical for endothelial cell-smooth muscle cell communication in venous morphogenesis.

Principle of the Assay  

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-TIE2 polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti-TIE2 polyclonal antibody was used as detection antibodies. The standards, test samples and biotin conjugated detection antibody were added to the wells subsequently, and wash with wash buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with wash buffer. TMB substrates were used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the TIE2 amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of TIE2 can be calculated.

Kit components

1. One 96-well plate pre-coated with anti-Human TIE2 antibody
2. Lyophilized Human TIE2 standards: 2 tubes (10 ng / tube)
3. Sample / Standard diluent buffer: 30ml
4. Biotin conjugated anti-Human TIE2 antibody (Concentrated): 130μl. Dilution: 1:100
5. Antibody diluent buffer: 12ml
6. Avidin-Biotin-Peroxidase Complex (ABC) (Concentrated): 130μl. Dilution: 1:100
7. ABC diluent buffer: 12ml
8. TMB substrate: 10ml
9. Stop solution: 10ml
10. Wash buffer(25X): 30ml

Note: Reconstitute standards and test samples with Kit Component 3.

Material Required But Not Provided

1. 37℃ incubator
2. Microplate reader (wavelength: 450nm)
3. Precise pipette and disposable pipette tips
4. Automated plate washer
5. ELISA shaker
6. 1.5ml of Eppendorf tubes
7. Plate cover
8. Absorbent filter papers
9. Plastic or glass container with volume of above 1L