Human uPAR ELISA Kit

Catalog No.: BEK1223

Size: 96T

Range: 62.5 pg/ml-4000 pg/ml

Sensitivity < 4 pg/ml

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

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

Introduction

The Urokinase receptor, also known as uPA receptor or uPAR or CD87 (Cluster of Differentiation 87), is multidomain glycoprotein tethered to the cell membrane with a glycosylphosphotidylinositol (GPI) anchor. uPAR was originally identified as a saturable binding site for urokinase on the cell surface. It consists of three different domains of the Ly-6/uPAR/ alpha-neurotoxin family. All three domains are necessary for high affinity binding of the primary ligand, urokinase. uPAR is a part of the plasminogen activation system, which in the healthy body is involved in tissue reorganization events such as mammary gland involution and wound healing. It is a key molecule in the regulation of cell-surface plasminogen activation, and plays an important role in many normal as well as pathologic processes.

Principle of the Assay  

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

Kit components

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