ATP levels were determined using an ATP Bioluminesence Assay Kit (Roche) as our previously described [7 ; 19 (link)]. Briefly, SK-N-SH cells were incubated with ABAD inhibitor and oligomer Aβ1-42 at 37°C for 48 hours. Cells were harvested using ATP lysis buffer followed by incubation for 30 minutes on ice. The mixture was centrifuged at 12,000 × g for 10 minutes at 4°C, and the supernatant was collected for the assay. The content of ATP was measured according to the manufacturer's instructions [3 (link); 7 ]. Light emitted from the luciferase-mediated reaction was captured in a luminescence plate reader (Molecular Devices) at 37°C with an integration time of 10 seconds and calculated from a log-log plot of the standard curve of known ATP concentrations.
Atp bioluminesence assay kit
Manufactured by Roche
The ATP Bioluminesence Assay Kit is a laboratory instrument designed to measure the amount of adenosine triphosphate (ATP) present in a sample. It utilizes bioluminescence technology to quantify the ATP levels, providing a rapid and sensitive method for various research and analytical applications.
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2 protocols using atp bioluminesence assay kit
1
Measuring ATP Levels in SK-N-SH Cells
2
ATP and ROS Quantification in Transgenic Mice
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ATP levels in brains of Tg mice were determined using an ATP Bioluminesence Assay Kit (Roche) following the manufacturer's instruction. Brain tissues were homogenized in the lysis buffer provided in the kit, incubated on ice for 15 min, and centrifuged at 14,000 g for 15 min. Subsequent supernatants were measured for the ATP levels using Luminescence plate reader (Molecular Devices) with an integration time of 10 seconds.
Evaluation of intracellular ROS levels was accessed by election paramagnetic resonance (EPR) spectroscopy as described in our previous study (Du et al.,2017 ). CMH (cyclic hydroxylamine 1‐hydroxy‐3‐methoxycarbonyl‐2,2,5,5‐tetramethyl‐pyrrolidine, 100 μM) was incubated with hippocampal slices for 30 min and then washed with cold PBS. The tissues were collected and homogenized with 100 μl of PBS for EPR measurement. The EPR spectra were collected, stored, and analyzed with a Bruker EleXsys 540x‐band EPR spectrometer (Billerica, MA) using the Bruker Software Xepr (Billerica, MA).
Evaluation of intracellular ROS levels was accessed by election paramagnetic resonance (EPR) spectroscopy as described in our previous study (Du et al.,
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