ELISA Kit for Prothrombin Fragment 1+2 (F1+2) Sus scrofa; Porcine (Pig) Sandwich ELISA

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  • ELISA Kit for Prothrombin Fragment 1+2 (F1+2) Packages (Simulation)
  • ELISA Kit for Prothrombin Fragment 1+2 (F1+2) Packages (Simulation)
  • ELISA Kit for Prothrombin Fragment 1+2 (F1+2) Results demonstration
  • SEA710Po.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant Prothrombin Fragment 1+2 (F1+2) and the recovery rates were calculated by comparing the measured value to the expected amount of Prothrombin Fragment 1+2 (F1+2) in samples.

Matrix Recovery range (%) Average(%)
sodium citrate plasma(n=5) 80-91 86

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Prothrombin Fragment 1+2 (F1+2) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Prothrombin Fragment 1+2 (F1+2) 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 Prothrombin Fragment 1+2 (F1+2) 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
sodium citrate plasma(n=5) 92-99% 94-102% 90-98% 80-93%

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.

ELISA Kit for Prothrombin Fragment 1+2 (F1+2)

Test principle

The microplate provided in this kit has been pre-coated with an antibody specific to F1. Standards or samples are then added to the appropriate microplate wells with a biotin-conjugated antibody specific to F2. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain F1+2, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of F1+2 in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • Disseminated intravascular coagulation or acute coagulopathy of trauma shock early after trauma? An observational studyBioMed: cc10553
  • Changes in thrombin generation, fibrinolysis, platelet and endothelial cell activity, and inflammation following endovascular abdominal aortic aneurysm repairScienceDirect: S0741521411018568
  • High levels of soluble VEGF receptor 1 early after trauma are associated with shock, sympathoadrenal activation, glycocalyx degradation and inflammation in severely injured patients: a prospective studySjtrem:1757-7241
  • High sCD40L levels early after trauma are associated with enhanced shock, sympathoadrenal activation, tissue and endothelial damage, coagulopathy and mortalityWiley: source
  • Evaluation of autologous plasma skin test in patients with chronic idiopathic urticariaPubMed: 21910697
  • Diet Modulates Endogenous Thrombin Generation, A Biological Estimate of Thrombosis Risk, Independently of the Metabolic StatusPubMed: 22859493
  • Medium-term effect of endovascular and open abdominal aortic aneurysm repair on thrombin generation and fibrinolysisPubMed: 23140799
  • Markers of Thrombogenesis and Fibrinolysis and Their Relation to Inflammation and Endothelial Activation in Patients with Idiopathic Pulmonary Arterial HypertensionPlosone: Source
  • Effect of endovascular and open abdominal aortic aneurysm repair on thrombin generation and fibrinolysisPubmed: 23140799
  • Clinical impact of factor V Leiden, prothrombin G20210A, and MTHFR C677T mutations among sickle cell disease patients of Central IndiaPubmed: 23992124
  • Study the association of adiponectin with inflammation and hypercoagulability in case of type 2 diabetic subjects with renal dysfunctionTandfonline:Source
  • An ex vivo evaluation of blood coagulation and thromboresistance of two extracorporeal circuit coatings with reduced and full heparin dosePubmed:24632424
  • Ex vivo simulation of cardiopulmonary bypass with human blood for hemocompatibility testingPubMed: 26243277
  • Comparison of the Effect of Dabigatran and Dalteparin on Thrombus Stability in a Murine Model of Venous ThromboembolismPubMed: 26514101
  • Thrombin activatable fibrinolysis inhibitor (TAFI), tissue factor pathway inhibitor (TFPI), and prothrombin fragment 1+2 levels in patients with advanced colorectal cancer aot:AOT_49_1_6_12.pdf
  • Perioperative thrombocytopenia predicts poor outcome in patients undergoing transcatheter aortic valve implantationpubmed:29145170
  • The causes of thrombocytopenia after transcatheter aortic valve implantationpubmed:28582640
  • Elevations of Thrombotic Biomarkers in Hemoglobin H DiseasePubmed:29402840
  • The cerebral thrombin system is activated after intracerebral hemorrhage and contributes to secondary lesion growth and poor neurological outcome in C57Bl/6 micePubmed: 31830857
  • Enhanced Fibrin-Lysis in Grade-1 Dengue Haemorrhagic Fever
  • The impact of anti-endothelial cell antibodies (AECAs) on the development of blood vessel damage in patients with systemic lupus erythematosus: the preliminary …Pubmed:35284968

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