Amplite® Colorimetric Malondialdehyde (MDA) Quantitation Kit

Protocol summary

1. Prepare test samples along with serially diluted MDA standards (50 µL)
2. Add MDA Blue™ stock solution (10 µL)
3. Incubate at room temperature for 10 - 30 minutes
4. Add Reaction Solution (40 µL)
5. Monitor OD increase at 695 nm

Important notes
Thaw all the components at room temperature before starting the experiment.

1. MDA standard solution (100 mM):
Add 100 µL of ddH₂O into MDA Standard vial (Component C) to make 100 mM MDA stock solution.

Table 1. Layout of MDA standards and test samples in a 96-well clear bottom microplate. MDA= MDA Standard (MDA1 - MDA7, 6.25 to 400 µM), BL=Blank Control, TS=Test Sample.

Table 2. Reagent composition for each well.

1. Prepare MDA standards (MDA), blank controls (BL), and test samples (TS) according to the layout provided in Tables 1 and 2. For a 384-well plate, use 25 µL of reagent per well instead of 50 µL.
   
   
2. Add 10 µL MDA Blue™ (Component A) solution into each well of MDA standard, blank control, and test samples. For a 384-well plate, use 5 µL MDA Blue™ solution into each well.
   
   
3. Incubate the reaction mixture at room temperature for 10 - 30 minutes.
   
   
4. Add 40 µL of Reaction Solution (Component D) to make the total assay volume of 100 µL/well. For a 384-well plate, use 20 µL of Reaction Solution, for a total assay volume of 50 µL/well.
   
   
5. Incubate the final reaction mixture at room temperature for 30 - 60 minutes.
   
   
6. Monitor absorbance increase with an absorbance plate reader with path-check correction at OD of 695 ~ 700 nm.