Question (75312) | ADP/ATP Ratio Assay Kit (Bioluminescent) (ab65313)

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Question

I would like to investigate the ratio of ATP:ADP in nerve tissue, namely DRG and saphenous nerve. Could you please advise whether this kit would be suitable and if extra steps are required?

Answer



Please see the following publication which may be of use to you:

Diguet N et al. Muscle creatine kinase deficiency triggers both actin depolymerization and desmin disorganization by advanced glycation end products in dilated cardiomyopathy. J Biol Chem 286:35007-19 (2011).

Protocol:
ATP and ADP/ATP ratios were measured using the bioluminescent ADP/ATP ratio assay kit from Abcam as indicated by the manufacturer with minor modifications. Briefly, cardiac tissue was frozen in liquid nitrogen and powdered with a mortar. Tissue powder was suspended in the provided lysis buffer (10 μl/mg of tissue powder) for 5 min exactly at room temperature, followed by centrifugation at 10,000 × g for 1 min to pellet insoluble material. We used 50 μl of supernatant per assay in a final volume of 100 μl. Luciferase units were converted to picomoles of the amount of ATP by plotting against a standard curve with serial dilution of ATP. Data were normalized by the amount of protein present in the supernatant as measured by the Bradford assay.

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