SPECTROZYME® FVIIa, chromogenic substrate for FVIIa assays
Chromogenic substrate for amidolytic assay of Factor VIIa and lipidated Tissue Factor/VIIa complex.
Chemical name: Methanesulphonyl-D-cyclohexyl-alanyl-butyl-arginine-paranitroaniline monoacetate salt
MW: 670.8 Dalton
Solubility: > 10 mM in distilled/deionized water,
> 0.5 mM in Tris Buffer, pH 8.4
SPECTROZYME® FVIIa, it is the substrate of choice in all chromogenic based methods for the determination of factor VIIa, and for the factor VIIa/Tissue Factor complex in particular.
Enzyme activity is determined by measuring the increase in absorbance of the free chromophore (pNA) generated, in comparison to the original substrate, per unit time at λ405 nm. At excess substrate concentration, the rate of absorbance increase due to the amount of chromophore released is linearly related to enzyme concentration. Measurement can be made either through acid quenching of the reaction (end-point method), or through use of a kinetic recording spectrophotometer (initial-rate-method).
Kinetic Data for Factor VIIa and TF complex
Buffer: 880 µL Tris-Buffer, pH 8.4 Temp. 37°C
(0.1M Tris, 0.1M NaCl, 5 mM CaCl2, 0.1% BSA)
Substrate: 0.100 mL of No. ADG217 at 4 mM
- human factor VIIa
0.020 mL of FVIIa at 0.025 mg/mL
=> KM : 5.0 mM Vmax: 6.72 nmoles/min
- By adding Tissue Factor with a 10:1 ratio (TF:FVIIa)
0.100 mL of FVIIa at 0.025 mg/mL +
0.100 mL of tissue factor at 0.25 mg/mL
=> KM : 0.97 mM Vmax: 69.7 nmoles/min
Glass vial containing 50 μmoles of lyophilized substrate.
Reconstitute with filtered distilled/deionized water to create a stock solution with a concentration of 5.0 mM. The typical working range concentration is 4.0 mM.
Store intact vials in the dark at 2 - 8 °C for up to 5 years from the date of manufacture. Reconsituted substrate may be stored for 1 week at room temperature, 2 months at 2° - 8°C, or for up to 6 months at –20°C (Aliquot and freeze. Do not submit to freeze-thaw cycles).
Category: Research use only
Product Availability: Worldwide
Manufacturer: ImmBioMed GmbH & Co KG, Germany
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SPECTROZYME FVIIa, chromogenic substrate for FVIIa assays
Einheit: 50 µmole
Hersteller: ImmBioMed GmbH & Co. KG