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ELISA 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) 95-105 101
EDTA plasma(n=5) 86-93 90
heparin plasma(n=5) 78-97 86

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) 89-103% 84-93% 80-96% 93-101%
EDTA plasma(n=5) 86-93% 80-93% 83-101% 79-98%
heparin plasma(n=5) 84-98% 93-101% 82-101% 93-101%

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
TMB Substrate 1×9mL Stop Solution 1×6mL
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 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

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Magazine Reference
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
Catalog No. Related products for research use of Mus musculus (Mouse) Organism species Applications (RESEARCH USE ONLY!)
APA172Mu01 Active Platelet Factor 4 (PF4) Cell culture; Activity Assays.
RPA172Mu01 Recombinant Platelet Factor 4 (PF4) Positive Control; Immunogen; SDS-PAGE; WB.
RPA172Mu02 Recombinant Platelet Factor 4 (PF4) Positive Control; Immunogen; SDS-PAGE; WB.
PAA172Mu01 Polyclonal Antibody to Platelet Factor 4 (PF4) WB; IHC; ICC; IP.
PAA172Mu02 Polyclonal Antibody to Platelet Factor 4 (PF4) WB; IHC; ICC; IP.
LAA172Mu71 Biotin-Linked Polyclonal Antibody to Platelet Factor 4 (PF4) WB; IHC; ICC.
LAA172Mu81 FITC-Linked Polyclonal Antibody to Platelet Factor 4 (PF4) WB; IHC; ICC; IF.
SEA172Mu ELISA Kit for Platelet Factor 4 (PF4) Enzyme-linked immunosorbent assay for Antigen Detection.
SCA172Mu CLIA Kit for Platelet Factor 4 (PF4) Chemiluminescent immunoassay for Antigen Detection.
KSA172Mu01 ELISA Kit DIY Materials for Platelet Factor 4 (PF4) Main materials for "Do It (ELISA Kit) Yourself".