Human P-Selectin ELISA Kit

Catalog No.: BEK1189

Size: 96T

Range: 156 pg/ml-10,000 pg/ml

Sensitivity < 5 pg/ml

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

Application: For quantitative detection of P-Selectin in serum, plasma, body fluids, tissue lysates or cell culture supernates. 

Introduction

P-selectin, also called GMP-140, CD62, or selectin P, is a 140-kD adhesion molecule that in humans is encoded by the SELP gene. It is located on chromosome 1q21-q24, spans>50 kb and contains 17 exons in human. It is constitutively expressed on megakaryocytes (the precursor of platelets) and endothelial cells. P-selectin plays an essential role in the initial recruitment of leukocytes (white blood cells) to the site of injury during inflammation. Ligands for P-selectin on eosinophils and neutrophils are similar sialylated, protease-sensitive, endo-beta-galactosidase-resistant structures, clearly different than those reported for E-selectin, and suggest disparate roles for P-selectin and E-selectin during recruitment during inflammatory responses.

Principle of the Assay  

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

Kit components

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