Amplite® Colorimetric Lipase Activity Assay Kit

1. Prepare test samples and the lipase standards (50 μL).

2. Add the lipase working solution (50 µL).

3. Incubate at 37 °C for 10-30 minutes.

4. Measure the absorbance at 586 nm.

Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles

1. Reconstitute Lipase Standard (Component D) by adding 100 µL of ddH2O to achieve a concentration of 1 mg/mL. Mix thoroughly by pipetting up and down several times.

   Note: The Lipase Standard Stock Solution can be stored at -20 °C, protected from light and should be used within 1 month after reconstitution.

    

1. Add 200 µL of Lipase Substrate (Component C) to 5 mL of Lipase Substrate Buffer (Component B). This 5 mL solution is sufficient for 100 tests. Prepare the required amount of Lipase Working Solution proportionally based on the number of tests you need.

1. Tissues and cells can be homogenized in the Lipase Assay Buffer (Component A). To remove insoluble material, centrifuge the sample at 13,000xg for 10 minutes. Serum samples can be added directly to the wells. Adjust the samples to a final volume of 50 µL using Lipase Assay Buffer.

Table 1. Layout of lipase standards and test samples in a 96-well clear bottom microplate. (STD = Lipase Standards (STD1-STD7, 0.625~40 µg/ml), BL= Blank Control, TS=Test Samples)

Table 2. Reagent composition for each well.

1. Prepare lipase standards (STD1-7), blank controls (BL), and test samples (TS) as outlined in Tables 1 and 2. For a 384-well plate, use 25 µL of reagent per well.

2. Add 50 µL of Lipase Working Solution to each well containing the blank control, Lipase Standards, and test samples (TS). If using a 384-well plate, add 25 µL of Lipase Working Solution to each well instead.

3. Incubate at 37 °C for 10-30 minutes, protected from light.

4. Monitor the absorbance at 586 nm.