Adenosine Diphosphate (ADP) is important compound found in living cells which can be phosphorylated into adenosine triphosphate (ATP) for energy storage and hydrolyzed back into ADP to release energy. ADP drives platelet aggregation and activation which is crucial in hemostasis. Dysfunction of ADP can lead to thrombosis, pulmonary embolism, and neurodegenerative disorders like Alzheimer’s. Monitoring ADP levels is important in understanding cellular energy and pathological conditions.

ADP Assay Kit provides a simple, reproducible, and sensitive tool of measuring ADP concentration in plasma, serum, cell lysates and other biological liquid samples. This assay employs a coupled enzymatic reaction system that combines multiple steps. Initially, pyruvate kinase catalyzes the conversion of ADP into ATP and pyruvate. Pyruvate is then oxidized with the conversion of the dye into a colored product that is measurable at 570 nm. The optical density measured at 570 nm is directly proportional to the concentration of ADP present in the sample.

• Microplate reader capable of measuring absorbance at 570 nm
• Precision pipettes to deliver 2 µL to 1 mL volumes
• Tubes to prepare sample dilutions
• 5 mL tubes
• MilliQ water or another type of double distilled water (ddH₂O)
• 10 kDa molecular weight cutoff (MWCO) centrifuge spin filter

1. Add 50 µL of the diluted ADP Standards or samples to the 96-well microtiter plate.
2. Add 50 µL Reaction Mix to each standard and one half of the paired sample wells, mix thoroughly.
3. Add 50 µL of Control Mix to other half of the paired sample wells. Carefully shake the plate for a few seconds to mix.
4. Seal the plate with a plate sealer or plastic wrap to reduce evaporation. Incubate the plate for 30 minutes at room temperature with mixing in the dark.
5. Measure the absorbance at 570 nm using a plate reader.