Human Angiopoietin-2, ANGPT2 ELISA Kit

Catalog No.: BEK1008

Size: 96T

Range: 8 pg/ml-150 pg/ml

Storage and Expiration: Store at 2-8℃ for 6 months. 

Application: For quantitative detection of ANGPT2 in Human serum, plasma, urine, cell culture supernatant or tissue samples.

Introduction

Angiopoietins are protein growth factors that promote angiogenesis, the formation of blood vessels from pre-existing blood vessels. All angiopoietins bind with similar affinity to an endothelial cell-specific tyrosine-protein kinase receptor. ANGⅡ is a naturally occurring antagonist of angiopoietin-1 (ANGPT1) that competes for binding to the TIE2 receptor and blocks ANGPT1-induced TIE2 autophosphorylation during vasculogenesis. ANGⅡ is a mediator of epithelial necrosis with an important role in hyperoxic acute lung injury and pulmonary edema. It plays a critical role in inducing tumor cell infiltration, and that this invasive phenotype is caused by activation of MMP2.

Principle of the Assay  

This kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. The purified anti-ANGPT2antibody was pre-coated onto 96-well plates. And the HRP conjugated anti-ANGPT2 antibody was used as detection antibodies. The standards, test samples and HRP conjugated detection antibody were added to the wells subsequently, mixed and incubated, then,

unbound conjugates were washed away with wash buffer. TMB substrates (A & B) 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 ANGPT2 amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of ANGPT2 can be calculated.

Kit components

1. One 96-well plate pre-coated with anti-human ANGPT2 antibody

2. Standard: 0.5ml (180 pg /mL)

3. Standard diluent buffer:  1.5 ml

4. Wash buffer (30×): 20 ml. Dilution: 1:30

5. Sample diluent buffer: 6 ml

6. HRP conjugated anti-human ANGPT2 antibody (RTU): 6ml

7. Stop solution: 6 ml

8. TMB substrate A:  6ml

9. TMB substrate B:  6ml

10. Plate sealer:   2

11. Hermetic bag:  1

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. Absorbent filter papers
8. Plastic or glass container with volume of above 1L