Chromogenic Substrate
Preparation
25 mg lyophilized FXa chromogenic substrate with mannitol added as a bulking agent. Reconstitute substrate with 20 ml sterile water.
Storage & Stability
The reconstituted substrate is stable for 6 months at 2 – 8°C.
Russell’s Viper Venom
Preparation
FX activating protein from Russell’s Viper Venom. Reconstitute the RVV with 15 ml sterile water.
Storage & Stability
The reconstituted activator is stable for 1 month at 2 – 8°C.
CaCl2
Preparation
20 ml of 0.1 mol/L calcium chloride solution. Before use, mix 1 volume of RVV with 1 volume of CaCl2. The mixture is stable for 48 hours at 2 – 8°C.
Storage & Stability
The solution is stable at 2 – 8°C until the expiry date printed on the label.
Buffer
Preparation
100 ml buffer solution containing 0.05 mol/L Tris, pH 7.8 and 20 mg/L Polybrene® (hexadimethrine bromide). Ready for use.
Storage & Stability
The buffer is stable at 2 – 8°C until the expiry date printed on the label.
The sealed reagents are stable at 2 – 8°C until the expiry date printed on the label. Contamination by microorganisms should be avoided once the vials are opened.
Materials required but not provided
Sterile water
Acetic acid, 20% (acid-stopped method only)
Spectrophotometer, 405 nm
Microplates if needed
Microplate reader if needed
Incubator 37° ± 0.2°C
Vortex mixer
Centrifuge, 2000 x g
Calibrated pipettes
Calibration plasma
Control plasma