ELISA Kit for Coagulation Factor V (F5) Homo sapiens (Human) Sandwich ELISA

FV; FVL; PCCF; Proaccelerin; Labile Factor; Factor V Leiden; Accelerator Globulin; Activated protein C cofactor; Proaccelerin, labile factor

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  • ELISA Kit for Coagulation Factor V (F5) Packages (Simulation)
  • ELISA Kit for Coagulation Factor V (F5) Packages (Simulation)
  • ELISA Kit for Coagulation Factor V (F5) Results demonstration
  • SEA675Hu.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant Coagulation Factor V (F5) and the recovery rates were calculated by comparing the measured value to the expected amount of Coagulation Factor V (F5) in samples.

Matrix Recovery range (%) Average(%)
sodium citrate plasma(n=5) 93-105 101

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Coagulation Factor V (F5) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Coagulation Factor V (F5) 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 Coagulation Factor V (F5) 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) 87-95% 94-103% 93-105% 98-105%

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
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 Coagulation Factor V (F5)

Test principle

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Coagulation Factor V (F5). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Coagulation Factor V (F5). 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 Coagulation Factor V (F5), 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 Coagulation Factor V (F5) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • Thrombin generation in abdominal sepsis is Rho-kinase-dependentPubMed: 25813486
  • Evidence of a Pivotal Role for the Distal Part of the Complement Cascade in the Diurnal Release of Hematopoietic Stem Cells Into Peripheral BloodPubmed:26087465
  • Loss of accelerates arterial thrombosis by increasing formation of neutrophil extracellular traps and plasma tissue factor activityPubmed:29444200

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