Human PF4 / CXCL4 ELISA Kit

Catalog No.: BEK1330

Size: 96T

Range: 156 pg/ml - 10,000 pg/ml

Sensitivity < 20 pg/ml

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

Application: For quantitative detection of PF4 / CXCL4 in human serum, plasma, body fluids, tissue lysates or cell culture supernatants.

Introduction

Platelet factor 4 (PF4) is a small cytokine belonging to the CXC chemokine family that is also known as chemokine (C-X-C motif) ligand 4 (CXCL4) . This chemokine is released from alpha-granules of activated platelets during platelet aggregation, and promotes blood coagulation by moderating the effects of heparin-like molecules. Due to these roles, it is predicted to play a role in wound repair and inflammation. It is usually found in a complex with proteoglycan. The gene for human PF4 is located on human chromosome 4.

Principle of the Assay  

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti- PF4 / CXCL4 polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti- PF4 / CXCL4 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 PF4 / CXCL4 amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of PF4 / CXCL4 can be calculated.

Kit components

1. One 96-well plate pre-coated with anti-human PF4 / CXCL4 antibody
2. Lyophilized human PF4 / CXCL4 standards: 2 tubes (50 ng / tube)                     
3. Sample / Standard diluent buffer: 30ml
4. Biotin conjugated anti-human PF4 / CXCL4 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