Mouse Angiopoietin-2, ANGPT2 ELISA Kit

Catalog No.: BEK1307

Size: 96T

Range: 156 pg/ml-10,000 pg/ml

Sensitivity < 10 pg/ml

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

Application: For quantitative detection of ANGPT2 in mouse serum, plasma, body fluids, tissue lysates or cell culture supernatants.

Introduction

Angiopoietin-2 is a protein that in humans is encoded by the ANGPT2 gene. Angiopoietins are members of the vascular endothelial growth factor family and the only known growth factors largely specific for vascular endothelium. ANGPT2 is a naturally occurring antagonist of angiopoietin-1 that competes for binding to the TIE2 receptor and blocks ANGPT1-induced TIE2 autophosphorylation during vasculogenesis. Its expression occurs at sites of vascular remodelling: dorsal aorta and major aortic branches, ovary, placenta and uterus.

Principle of the Assay

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti- ANGPT2 polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti- ANGPT2 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 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-mouse ANGPT2 antibody
2. Lyophilized mouse ANGPT2 standards: 2 tubes (10ng / tube)
3. Sample / Standard diluent buffer: 30ml
4. Biotin conjugated anti-mouse ANGPT2 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
11. 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