Human Heparin-platelets factor 4 antibody, HPF4-Ab ELISA Kit

Catalog No.: BZEK2420-48

Size: 48T

Range: 0.75 μg/L - 12 μg/L

Sensitivity: 0.1 μg/L

Storage and Expiration: Store at 2-8℃ for 6 months. 

Application: For quantitative detection of HPF4-Ab in human serum, plasma, tissue homogenate or cell culture supernatant.

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 heparin:PF4 complex is the antigen in heparin-induced thrombocytopenia (HIT), an idiosyncratic autoimmune reaction to the administration of the anticoagulant heparin. PF4 autoantibodies have also been found in patients with thrombosis and features resembling HIT but no prior administration of heparin.

Principle of the Assay  

This kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. The recombinant human HPF4 antigen was pre-coated onto 48-well plates. And the HRP conjugated anti- HPF4-Ab antibody was used as detection antibodies. The standards, test samples and HRP conjugated detection antibody were added to the wells subsequently, mixed and incubated, then, unbound conjugates were washed away with wash buffer. TMB substrates (A & B) 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 HPF4-Ab amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of HPF4-Ab can be calculated.

Kit components

1. One 48-well plate pre-coated with human HPF4 antigen

2. Standard: 0.5ml (24 μg/L)

3. Standard diluent buffer: 1.5 ml

4. Wash buffer (20×): 20 ml. Dilution: 1:20

5. Sample diluent buffer: 3 ml

6. HRP conjugated anti- human HPF4-Ab antibody (RTU): 3ml

7. Stop solution: 3 ml

8. TMB substrate A:  3ml

9. TMB substrate B:  3ml

10. Plate sealer:   2

11. Hermetic bag:  1

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. Absorbent filter papers
8. Plastic or glass container with volume of above 1L