Liver and tail tissues near the 6th caudal vertebrae were collected and weighed, after which ATP lysis buffer (100 μL/10 mg sample, S0026, Beyotime Biotechnology, Nantong, China) was added into the tubes and the tissues were minced. The samples were incubated on ice for 5 min and then centrifuged at 12,000 × g at 4 °C. The supernatant (50 μL) was transferred into an opaque 96-well plate, which was pre-loaded with 100 μL ATP detection buffer. The absorbance of the samples was measured with an illuminometer (Synergy TM 2, BioTek, Waldbronn, Germany).
S0026
Manufactured by Beyotime
Sourced in China
S0026 is a laboratory equipment product. It serves as a core function for laboratory applications. Further details on its intended use are not available.
Automatically generated - may contain errors
Lab products found in correlation
Synergy 2, by Agilent Technologies (3 mentions)
Bca protein assay kit, by Beyotime (2 mentions)
2 nbdg, by Cayman Chemical (2 mentions)
Fluorescence microplate reader, by Agilent Technologies (2 mentions)
Microporous plate photometer, by Agilent Technologies (2 mentions)
Microplate reader, by Agilent Technologies (2 mentions)
Tristar2 lb 942, by Berthold Technologies (1 mentions)
Atp lysis buffer, by Beyotime (1 mentions)
Bc1175, by Solarbio (1 mentions)
Glutathione (gsh), by Solarbio (1 mentions)
Bc0170, by Solarbio (1 mentions)
Sorvall legend micro 17r centrifuge, by Thermo Fisher Scientific (1 mentions)
Infinite 200 pro, by Tecan (1 mentions)
S0175, by Beyotime (1 mentions)
Spectramax l, by Molecular Devices (1 mentions)
Multifunctional microplate reader, by Tecan (1 mentions)
Varioskan lux, by Thermo Fisher Scientific (1 mentions)
Varioskan flash, by Thermo Fisher Scientific (1 mentions)
Microplate reader, by Molecular Devices (1 mentions)
Fluorescence multi detection microplate reader, by Agilent Technologies (1 mentions)
79 protocols using s0026
1
Tissue ATP Extraction and Quantification
2
ATP Quantification Using Luciferase Assay
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ATP content was measured using a firefly luciferase-based ATP assay kit (S0026, Beyotime) according to the manufacturer’s instructions. In brief, cells were lysed in 200 μl of lysis buffer/3 cm dish and collected for centrifugation at 12,000 × g for 5 min at 4 °C. The supernatants were collected. Two microliters were used for protein concentration measurement, and the remnant was incubated with the luciferin substrate and luciferase enzyme in the dark for 1 min to stabilize the luminescent signal. A fluorescence microplate reader was used to measure the bioluminescence intensity. The ATP concentration was calculated from the standard curve.
3
ATP Production Quantification
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The ATP amount produced in cells was assessed according to the standard protocol (Beyotime, S0026, Nanjing, China). Cells were lysed in lysis buffer, and then the supernatant was taken for subsequent determination after centrifugation for 5 min at 4 ℃. The supernatant was incubated for 5 min in a proportional ATP detection working solution at room temperature, followed by equal mixing with the standard solution and measurement of the RLU or CPM value with a chemiluminescence meter. The ATP value in OSKM-Wdr82 cells was obtained after normalization of the background and the subsequent OSKM control experiments.
4
Quantifying RhoA Activation and ATP
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Activated RhoA (GTP-bound) within cells was measured by a colorimetric method according to the product manual (Cytoskeleton, BK124). ATP content within cells was measured by a luciferase method according to the protocol of ATP detection kit (Beyotime, S0026).
5
Cellular ATP Quantification
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According to the manufacturer's instruction, an ATP content kit (S0026, Beyotime) was used to detect cellular ATP content. ATP level was further normalized to protein content.
6
Measuring ATP Levels in BV2 Cells
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ATP levels were measured using a firefly luciferase-based ATP detection kit (S0026, Beyotime, China). After being washed with PBS, BV2 cells were lysed using ATP detection lysis buffer, followed by centrifugation at 12 000 × g for 5 min at 4 °C, and the supernatant was collected. Then, 100 µL of the supernatant was mixed with 100 µL of ATP detection solution in a 1.5 mL tube. Luminescence (corresponding to total ATP levels) was immediately measured in relative light units (RLU) (nmol/mg) using a Turner Biosystems luminometer (Tecan, Switzerland). Finally, the ATP level of each sample was determined according to the RLU value of the standard sample and normalized to the protein concentration.
7
Leydig Cell ATP Quantification
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The ATP levels of Leydig cells were measured by using a firefly luciferase based on ATP assay kit (S0026, Beyotime) according to the manufacturer's instructions. The fluorescence luminance (RLU) was measured with the luminometer (Multimode Reader LB942 TriStar2, BERTHOLD, Germany). The unit of ATP was corrected to nmol/mg with concentration of mitochondrial protein.
8
Quantification of Cellular ATP Levels
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Macrophages from each group were collected and resuspended in PBS (1×106 cells/ml). ATP lysis buffer (Beyotime Institute of Biotechnology Co., Ltd.) was added to the cells, then mixed and incubated for 30 min on ice. The cells were centrifuged at 12,000 × g for 5 min at 4°C and then the ATP level in the supernatant was measured according to the specific steps of the ATP assay kit (S0026, Beyotime Institute of Biotechnology Co., Ltd.). The luminescence signals were measured using a BCA protein assay kit (P0010S, Beyotime Institute of Biotechnology Co., Ltd.) and a BMG reader (Thermo Fisher Scientific, Inc.). Finally, the ATP concentration was normalized to the protein concentration against ATP standard solution (Beyotime Institute of Biotechnology Co., Ltd.).
9
Mitochondrial Function and Oxidative Stress
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10
ATP Quantification in Cell Lysates
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Intracellular ATP contents were detected using an ATP assay kit (S0026; Beyotime)
following the manufacturer’s protocol. Harvested cells were lysed using the
lysis buffer and centrifuged at 12,000 g for 5 min at 4°C. The supernatants were
then harvested, and ATP was detected using a luminometer. The concentration of
ATP in each sample was calculated according to a standard curve and normalized
using the cellular protein level.
following the manufacturer’s protocol. Harvested cells were lysed using the
lysis buffer and centrifuged at 12,000 g for 5 min at 4°C. The supernatants were
then harvested, and ATP was detected using a luminometer. The concentration of
ATP in each sample was calculated according to a standard curve and normalized
using the cellular protein level.
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