Human PAI-1 ELISA Kit

Catalog No.: BEK1184

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 PAI-1 in plasma, tissue lysates or cell culture supernatants.

Introduction

Plasminogen activator inhibitor-1 (PAI-1) also known as endothelial plasminogen activator inhibitor or serpin E1 is a protein that in humans is encoded by the SERPINE1 gene. This gene is approximately 12.2 kb long and contains 9 exons, and located on the seventh chromosome (7q21.3-q22). PAI-1 is a member of the serine protease inhibitor family that inhibits tissue-type plasminogen activator and urokinase-type plasminogen activator. PAI-1 is mainly produced by the endothelium (cells lining blood vessels), but is also secreted by other tissue types, such as adipose tissue. PAI-1 is present in increased levels in various disease states (such as a number of forms of cancer), as well as in obesity and the metabolic syndrome. It has been linked to the increased occurrence of thrombosis in patients with these conditions.

Principle of the Assay  

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-PAI-1 polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti-PAI-1 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 PAI-1 amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of PAI-1 can be calculated.

Kit components

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