Cardiomyocytes were washed twice with cold PBS and lysed with lysis buffer containing phosphatase inhibitors for 30 min at 4°C. Samples were centrifuged at 13,000 rpm for 15 min at 4°C, and the supernatant was collected for use in the firefly luciferase-based ATP assay according to the manufacturer’s instructions. The luminescence signal was measured using a luminometer (Tecan Infinite M200 Pro) at 560-nm absorbance. The firefly luciferase-based ATP determination kit (Molecular Probes) was used to measure mitochondrial ATP content. Each assay was performed at least three times (Yan et al., 2019 (link)).
Firefly luciferase based atp determination kit
Manufactured by Thermo Fisher Scientific
The Firefly luciferase-based ATP determination kit is a laboratory product that measures the concentration of adenosine triphosphate (ATP) in a sample. The kit utilizes the bioluminescent reaction catalyzed by firefly luciferase to quantify the amount of ATP present.
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2 protocols using firefly luciferase based atp determination kit
1
Measuring Mitochondrial ATP Content
2
Quantifying Cellular ATP Levels
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The cellular ATP content was measured with a firefly luciferase-based ATP determination kit (Molecular Probes). Briefly, cardiomyocytes were washed twice with cold PBS and then lysed with lysis buffer containing phosphatase inhibitors for 30 min at 4 °C. Samples were centrifuged at 13,000 rpm for 15 min at 4 °C, and the supernatant was collected for use in the firefly luciferase-based ATP assay according to the manufacturer’s protocol. The luminescence signal was measured by using a luminometer (TECAN Infinite M200 Pro) at 560-nm absorbance. For each assay, at least 3 replicates were used.
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