Published on 2014/6/5 17:06:09
In order to use heparinase I more precisely, customers are advised to take the following quick assay, which gives the activity of heparinase I on heparin.
1. Prepare a 1 mg/mL heparin solution with 50 mM Tris-HCl buffer containing 10 mM CaCl2, pH 7.0. Incubate at 35 °C in water bath before use.
2. Transfer 2.5 mL of heparin solution into a 5 mL quartz cuvette (1 cm optical path).
3. Transfer 5 to 20 uL of heparinase I solution into the aforementioned cuvette. Mix gently.
4. Use an UV spectrometer to measure the change of absorbance per minute at 232 nm.
5. Calculate the enzymatic activity in international unit (IU) by the following formula
Activity = (Change of absorbance per minute at 232 nm)/(Volume of enzyme used) * 714
Where the enzymatic activity is measured in IU/mL, the volume of enzyme used is measured in uL.
For example, the volume of enzyme you transfer into the cuvette is 20 uL, and the change of absorbance per minute at 232 nm is 0.34, then the activity is 0.34/20*714 IU/mL = 12.1 IU/mL.
Know more about heparinases.