Factor VII (FVII) is a glycoprotein synthesized by the liver, vitamin k dependent. When tissue factor (TF) appears on the surface of damaged, abnormal or activated vascular endothelium, FVIIa associates with it, initiating the pathway extrinsic coagulation.
The FT-FVIIa complex activates the FX in FXa and the FIX in FIXa.
- Stability 1 month after opening
- Reaction time 120 minutes
- This test recognizes both native and recombinant human FVIIa and FVIIa/TF complexes
- FVII is not detected in the test
- FVII does not auto-activate in FVIIa during the execution of this test
- FVIIa in normal plasmas is approximately 5 ng/mL
- Sensitivity between 0.6 to 100 ng/mL
The IMUBIND FVIIa ELISA assay uses a biotinylated FVIIa enzyme inhibitor and anti-FVII / FVIIa monoclonal antibody as the capture antibody.
Diluted plasma samples or supernatants containing FVIIa are incubated with the biotinylated inhibitor, which covalently binds to FVIIa but not FVII.
The samples are added to the microwell coated with the capture monoclonal antibody. The FVIIa is detected thanks to the streptavidin-HRP which will bind the FVIIa complex captured at the bottom of the well by the monoclonal antibody and the biotinylated FVIIa inhibitor.
The TMB will thus recognize the HRP giving a blue compound which will be stopped by adding sulfuric acid giving a yellow compound, measured at 450nm. The results will be compared with a known FVIIa standard curve.
- 12 x 8-well breakable ELISA strips coated with anti-human FVII / FVIIa monoclonal antibody
- 2 vials of FVIIa standard, 200 ng / mL lyophilized
- 1 vial of FVII deficient plasma, 0.5 mL lyophilized
- 1 vial of reference plasma, 300 µL lyophilized
- 1 vial of FVIIa inhibitor, biotinylated, 200 µL freeze-dried concentrate
- 1 vial of enzyme conjugate, streptavidin-HRP, 120 µL
- 1 vial of TMB substrate, 11 mL
- 1 vial of stabilizer, 4.0 mL lyophilized
- 1 vial of test diluent, 22 mL lyophilized
- 1 packet of wash buffer, PBS with Tween 20 0.05%