D-VLK Plasmin Substrate Protocol

1. Make a D-VLK stock solution of 10 mM in 1 mM HCl.
Molecular Weight = 551
For the 25 mg vial, add 4.5 ml 1 mM HCl directly to vial.
For the 100 mg vial, add 18 ml 1 mM HCl directly to vial.
Aliquot and freeze at -20 C or colder.

2. Make a D-VLK working dilution of 200 uM in 0.1 M Hepes; 0.1 M NaCl; pH 7.4.
Add 200 ul of 10 mM D-VLK to 10 ml of buffer.
Store working dilution at 4 C.
Warm working dilution to room temperature before use.

3. Pipette 900 ul into a disposable cuvette, add 10 ul of test sample and mix well.
Actual volumes will vary based on cuvette size, sample plasmin activity, and end users laboratory conditions.

4. Monitor change in absorbance at 405 nm for 1 minute.

5. Calculate rate based on slope of curve.

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