Magnetic Luminex Assay Kit for Platelet Factor 4 (PF4) ,etc.

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

(Note: Up to 8-plex in one testing reaction)

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 86-95 90
EDTA plasma(n=5) 93-101 96
heparin plasma(n=5) 87-101 92
sodium citrate plasma(n=5) 94-103 98

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Platelet Factor 4 (PF4) ,etc. 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) ,etc. 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) ,etc. 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) 95-103% 80-89% 78-94% 88-99%
EDTA plasma(n=5) 78-97% 84-99% 98-105% 87-98%
heparin plasma(n=5) 85-92% 83-92% 96-104% 78-96%
sodium citrate plasma(n=5) 78-97% 95-102% 84-95% 96-105%

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
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:PF4) 1 Analysis buffer 1×20mL
Pre-Mixed Detection Reagent A 1×120μL Assay Diluent A 1×12mL
Detection Reagent B (PE-SA) 1×120μL Assay Diluent B 1×12mL
Sheath Fluid 1×10mL Wash Buffer (30 × concentrate) 1×20mL
Instruction manual 1

Assay procedure summary

1. Preparation of standards, reagents and samples before the experiment;
2. Add 100μL standard or sample to each well,
    add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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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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