CLIA Kit for Platelet Factor 4 (PF4)

CXCL4; SCYB4; Chemokine C-X-C-Motif Ligand 4; Oncostatin-A; Iroplact

Specificity

This assay has high sensitivity and excellent specificity for detection of Platelet Factor 4 (PF4).
No significant cross-reactivity or interference between Platelet Factor 4 (PF4) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Platelet Factor 4 (PF4) and the recovery rates were calculated by comparing the measured value to the expected amount of Platelet Factor 4 (PF4) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 84-103 98
EDTA plasma(n=5) 91-104 101
heparin plasma(n=5) 93-102 96

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Platelet Factor 4 (PF4) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Platelet Factor 4 (PF4) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Platelet Factor 4 (PF4) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 86-95% 83-97% 84-92% 95-102%
EDTA plasma(n=5) 89-96% 89-96% 95-104% 86-104%
heparin plasma(n=5) 85-98% 89-96% 90-98% 94-103%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
Substrate A 1×10mL Substrate B 1×2mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 100µL Substrate Solution. Incubate 10 minutes at 37°C;
8. Read RLU value immediately.

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Magazine Citations
Biochimica et Biophysica Acta (BBA) - Proteins & Proteomics Propofol lowers serum PF4 level and partially corrects hypercoagulopathy in endotoxemic rats PubMed: 20601223
Basic Research in Cardiology Secretome of apoptotic peripheral blood cells (APOSEC) attenuates microvascular obstruction in a porcine closed chest reperfused acute myocardial infarction model: role of platelet aggregation and vasodilation PubMed: 22899170
PLoS One Platelets Recognize Brain-Specific Glycolipid Structures, Respond to Neurovascular Damage and Promote Neuroinflammation PubMed: PMC3608633
Diving and Hyperbaric Medicine Effect of splenectomy on platelet activation and decompression sickness outcome in a rat model of decompression Pubmed:25311322
Journal of Functional Foods A Phellinus baumii–based supplement containing Salvia miltiorrhiza Bunge improves atherothrombotic profiles through endothelial nitric oxide synthase and cyclooxygenase pathwaysin vitro and in vivo science:S1756464616300676
Plos one Aspirin, but Not Tirofiban Displays Protective Effects in Endotoxin Induced Lung Injury pubmed:27583400
international journal of molecular sciences Evidence of the Role of R-Spondin 1 and Its Receptor Lgr4 in the Transmission of Mechanical Stimuli to Biological Signals for Bone Formation. pubmed:28272338
Experimental and Therapeutic Medicine Feasibility of improving platelet‑rich plasma therapy by using chitosan with high platelet activation ability pubmed:28450960
Reproductive Toxicology Aspirin pre-treatment modulates ozone-induced fetal growth restriction and alterations in uterine blood flow in rats Pubmed: 30528429
INTERNATIONAL JOURNAL OF CARDIOLOGY Short and long-term changes in platelet and inflammatory biomarkers after cryoballoon and radiofrequency ablation Pubmed: 30857843
Available at SSRN 3335365  Evaluating the Platelet Activation Related to the Degradation of Biomaterials by Scheme of Molecular Markers
journal of biotechnology Newly Identified HNP‐F from Human Neutrophil Peptide 1 Promotes Hemostasis Pubmed: 30927490
Data in Brief Data for short and long-term prothrombotic biomarkers after cryoballoon and radiofrequency ablation
journal of thoracic and cardiovascular surgery Evaluating Platelet Activation Related to the Degradation of Biomaterials Using Molecular Markers Pubmed: 32812629
Marine Drugs The Inhibitory Effect of Protamine on Platelets is Attenuated by Heparin without Inducing Thrombocytopenia in Rodents Pubmed: 31533230
Thrombosis Research Feasibility study of use of rabbit blood to evaluate platelet activation by medical devices Pubmed: 31838449
Nat Commun The C5a/C5a receptor 1 axis controls tissue neovascularization through CXCL4 release from platelets 34099640
Vaccine The effect of ChAdOx1 nCov-19 vaccine on arterial thrombosis development and platelet aggregation in female rats Pubmed:35183388
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