Human ADAMTS13 ELISA Kit

Catalog No.: BEK1329

Size: 96T

Range: 156 pg/ml - 10,000 pg/ml

Sensitivity < 26 pg/ml

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

Application: For quantitative detection of ADAMTS13 in human serum, plasma, body fluids, tissue lysates or cell culture supernatants.

Introduction

ADAMTS13 (a disintegrin and metalloproteinase with a thrombospondin type 1 motif, member 13)—also known as von Willebrand factor-cleaving protease (VWFCP)—is a zinc-containing metalloprotease enzyme that cleaves von Willebrand factor (vWf), a large protein involved in blood clotting. It is secreted into the blood and degrades large vWf multimers, decreasing their activity, hence ADAMTS13 acts to reduce thrombus formation. The ADAMTS13 gene maps to the ninth chromosome (9q34).

Principle of the Assay  

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti- ADAMTS13 polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti- ADAMTS13 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 ADAMTS13 amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of ADAMTS13 can be calculated.

Kit components

1. One 96-well plate pre-coated with anti-human ADAMTS13 antibody
2. Lyophilized human ADAMTS13 standards: 2 tubes (50 ng / tube)                     
3. Sample / Standard diluent buffer: 30ml
4. Biotin conjugated anti-human ADAMTS13 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