Published on 2014/8/5 10:30:54
Take 18 microcentrifuge tubes (1.5 mL, with lids), label and arrange the tubes as B1, S1, S2, S3, S4, T1, T2, T3, T4, T1, T2, T3, T4, S1, S2, S3, S4, B2.
Add each reagent as Table 3.
|Sequence||Solution||Volume||Incubation Temperature||Reaction time|
|Note: After each addition of a reagent, the incubation mixture should be mixed without allowing bubbles to form.|
or Tris-NaCl buffer (pH 7.4)
|50 uL||37 °C||-|
|2||Antithrombin||50 uL||37 °C||1 minute|
|3||Thrombin||100 uL||37 °C||1 minute|
|4||Chromogenic substrate||250 uL||37 °C||4 minutes|
|5||Stop solution||375 uL||-||-|
Prepare a reference solution according to Table 3. Use S1 and don't add thrombin solution.
Measure the absorbance of each tube at 405 nm wavelength using a spectrophotometer.
Calculate the anti-FIIa potency with a parallel-line assay program.