To quantify the metabolic activity of Vorganoids formed at different stages. RIPA buffer was added to lyse the Vorganoids, which were subsequently centrifuged at 4 °C for 5 min at 12000 rpm, after which the supernatant was removed for adenosine triphosphate (ATP) determination. The ATP assay working solution was prepared according to the kit instructions (S0026, Beyotime, China), and an ATP standard curve was constructed. The ATP working solution (100 μL) was added to the test wells and allowed to stand for 3-5 min at room temperature to eliminate background effects. Then, an appropriate amount of sample or standard was added and mixed well. The relative light unit value was measured using a chemiluminescence metre.
Protocol detail
Quantifying Vorganoid Metabolic Activity
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Corresponding Organization :
Other organizations : Southern Medical University, Guangdong Provincial People's Hospital, Zhuhai People's Hospital, Jinan University, University of Macau
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Variable analysis
independent variables
- Stages of Vorganoid formation
- Metabolic activity of Vorganoids
- RIPA buffer used to lyse Vorganoids
- Centrifugation at 4 °C for 5 min at 12000 rpm
- ATP assay working solution preparation according to kit instructions
- ATP standard curve construction
- Incubation of ATP working solution with samples/standards for 3-5 min at room temperature
- ATP standard curve
- Not explicitly mentioned
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