Cell Meter™ Intracellular Colorimetric Lipid Peroxidation (MDA) Assay Kit

1. Prepare and add MDA standards and/or test samples (50 µL)
2. Prepare and add MDA Blue™ (10 µL)
3. Incubate at room temperature for 10 - 30 minutes
4. Add reaction solution (40 µL)
5. Monitor OD increase at 695 nm

Thaw all the kit components at room temperature before starting the experiment.

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

Add 100 µL of ddH₂O into MDA Standard (Component C) and mix them well. Note: The unused MDA stock solution should be stored at -20 °C in single use aliquots.

Table 1. Layout of MDA standards and test samples in a clear bottom 96-well microplate. MDA = MDA standard (MDA1-MDA7= 1000 to 50 µM); BL = blank control; TS = test sample.

Table 2. Reagent composition for each well.

1. Add 50 µL of MDA standard, blank control, and test samples to clear bottom 96-well microplate (As shown in Table 1 and Table 2).

2. Add 10 µL/well of MDA Blue™ (Component A) into each well of MDA standard, blank control and test samples. Note: For a 384-well plate, add 25 µL of sample, 5 µL of MDA Blue™ solution into each well. Note: Please aliquot Component A into single use size and store unused at -20 ^oC and avoid light.

3. Incubate the reaction at room temperature for 10 - 30 minutes, protected from light.

4. Add 40 µL of Reaction Solution (Component D) to make the total assay volume of 100 µL/well. Note: For a 384-well plate, add 20 µL of Reaction Solution (Component D) to make the total assay volume of 50 µL/well.

5. Monitor absorbance increase with an absorbance plate reader with path-check correction at OD of 695~700 nm.