Human Angiopoietin-1, ANGPT1 ELISA Kit

Catalog No.: BEK1278

Size: 96T

Range: 156 pg/ml - 10,000 pg/ml

Sensitivity < 12 pg/ml

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

Application: For quantitative detection of ANGPT1 in human serum, plasma, body fluids, tissue lysates or cell culture supernatants.

Introduction

Angiopoietin 1, also called ANG1 is a type of angiopoietin and is encoded by the gene ANGPT1. Angiopoietins are proteins with important roles in vascular development and angiogenesis. All angiopoietins bind with similar affinity to an endothelial cell-specific tyrosine-protein kinase receptor. This gene was mapped to 8q23.1. The protein encoded by this gene is a secreted glycoprotein that activates the receptor by inducing its tyrosine phosphorylation. It plays a critical role in mediating reciprocal interactions between the endothelium and surrounding matrix and mesenchyme and inhibits endothelial permeability. The protein also contributes to blood vessel maturation and stability, and may be involved in early development of the heart.

Principle of the Assay

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti- ANGPT1 polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti- ANGPT1 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 ANGPT1 amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of ANGPT1 can be calculated.

Kit components

1. One 96-well plate pre-coated with anti-human ANGPT1 antibody
2. Lyophilized human ANGPT1 standards: 2 tubes (10ng / tube)                     
3. Sample / Standard diluent buffer: 30ml
4. Biotin conjugated anti-human ANGPT1 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