A095 1 1

Manufactured by Nanjing Jiancheng

Sourced in China

The A095-1-1 is a laboratory equipment item. It is designed for laboratory use, but its core function is not elaborated on further in order to maintain an unbiased and factual approach.

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Close spelling variants of this product

A095-1 (2 citations) ATP (A095-1-1) (2 citations) ATP assay kit (A095-1-1) (2 citations)

17 protocols using a095 1 1

Fly Metabolism and Antioxidant Assays

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The flies were crossed on standard food or RJ food. A few days later, hatched adult flies from standard food or RJ food were collected and cultured on standard food or RJ food. The 7-day-old male flies were homogenized in a buffer solution of phosphate-buffered saline (PBS) and then centrifuged for 10 min at 13,000 g. The supernatant was transferred to a new tube, the MDA (A003-1, Jiancheng Bio-Engineering, Nanjing, China), SOD oxidase (A001-3, Jiancheng Bio-Engineering, Nanjing, China), CAT oxidase (A007-1, Jiancheng Bio-Engineering, Nanjing, China), triglyceride (F001-1, Jiancheng Bio-Engineering, Nanjing, China), and ATP (A095-1-1, Jiancheng Bio-Engineering, Nanjing, China) kits were used to measure the amounts of components, according to the manufacturer's instructions. The total protein was measured using the BCA Assay Kit (CW0014, CoWin Biosciences, Beijing, China) and used for normalization. The resulting data were analyzed using one-way ANOVA.

Cheng Y., Cai J., Fu Y., Feng C., Hao Y, & Wei Y. (2020). Royal jelly attenuates metabolic defects in a Drosophila mutant with elevated TORC1 activity. Biology Open, 9(11), bio054999.

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Monascus Mycelium Enzyme Activities

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SBF broth at 84 h and 168 h were centrifuged at 8085× g and at 4 °C for 10 min to collect Monascus mycelium, then 0.1 g of the collected mycelium were broken in nine-fold volume of ice-cold phosphate buffered solution (PBS) (50 mM, pH = 6.8) to generate the mycelium lysate. Then, the lysate was centrifuged at 8085× g and at 4 °C for 10 min to obtain the supernatant. The activities of peroxidase (POD, EC:1.11.1.7), total kinds of superoxide dismutase (T-SOD, EC1.15.1.1), ETC complex I, II and III IV were determined by using the assay kits (Nanjing Jiancheng Bioengineering Institute, Nanjing, China) [20 (link)]. Protein concentrations were measured using the conventional Bradford assay. Reactive oxygen species (ROS) was measured according to the protocol by using the assay kit (E004-1-1, Nanjing Jiancheng Bioengineering Institute, Nanjing, China). The concentrations of ATP were detected by using the assay kit (A095-1-1, Nanjing Jiancheng Bioengineering Institute, Nanjing, China).

Cai X., Zhang S., Lin J., Wang Y., Ye F., Zhou B., Lin Q, & Liu J. (2022). Role of the Gene ndufs8 Located in Respiratory Complex I from Monascus purpureus in the Cell Growth and Secondary Metabolites Biosynthesis. Journal of Fungi, 8(7), 655.

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Oocyte ATP Quantification Protocol

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The ATP levels in the oocytes were detected using a commercial ATP assay kit (A095-1-1, Nanjing Jiancheng, Nanjing, China) according to the manufacturer’s instructions. Briefly, 300 μL of hot distilled water containing 60–80 oocytes were transferred into a glass homogenizer and homogenized. The resulting cell suspension was heated in a boiling water bath for 10 min, vortexed, and mixed for 1 min. A colorimetric method was immediately used to determine ATP concentrations using a spectrophotometer (Unico 7200, Franksville, WI, USA). The calculation method of the ATP concentrations in each oocyte (pmol/oocyte) was as described previously [40 (link)]. The ATP production was detected three times.

Lan T., Zhang K., Lin F., He Q., Wu S., Xu Z., Zhang Y, & Quan F. (2022). Effects of MICU1-Mediated Mitochondrial Calcium Uptake on Energy Metabolism and Quality of Vitrified-Thawed Mouse Metaphase II Oocytes. International Journal of Molecular Sciences, 23(15), 8629.

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Determination of Muscle ATP Content

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The ATP content in muscle tissues and lysed cells were determined via phosphomolybdic acid colorimetric method according to the manufacturer’s instructions (A095-1-1, Nanjing Jiancheng Bioengineering Institute, Nanjing, China).

You B., Dun Y., Fu S., Qi D., Zhang W., Liu Y., Qiu L., Xie M, & Liu S. (2021). The Treatment of Rhodiola Mimics Exercise to Resist High-Fat Diet-Induced Muscle Dysfunction via Sirtuin1-Dependent Mechanisms. Frontiers in Pharmacology, 12, 646489.

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Muscle Metabolic Enzyme Quantification

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The adenosine 5′-triphosphate (ATP), lactate dehydrogenase (LDH) and succinate dehydrogenase (SDH) contents in mucles were determined in accordance with the manufacturer’s procedures (A095-1-1, A020-2, A022-1-1, Nanjing Jiancheng) by a spectrophotometer and microplate reader.

Li Q., Wu J., Huang J., Hu R., You H., Liu L., Wang D, & Wei L. (2022). Paeoniflorin Ameliorates Skeletal Muscle Atrophy in Chronic Kidney Disease via AMPK/SIRT1/PGC-1α-Mediated Oxidative Stress and Mitochondrial Dysfunction. Frontiers in Pharmacology, 13, 859723.

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ATP and ROS Quantification in Breast Muscle

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The ATP levels in each breast muscle sample were determined by using a commercial assay kit (no. A095-1-1, Nanjing Jiancheng Bioengineering Institute, Nanjing, China) according to the manufacturer's instructions. The concentration of ROS in breast muscle was measured using an ELISA kit (no. MM-6012001, Jiangsu Meimian Industrial Co., Ltd., Jiangsu, China) according to the manufacturer's instructions.

Jing J., Wang J., Xiang X., Yin S., Tang J., Wang L., Jia G., Liu G., Chen X., Tian G., Cai J., Kang B., Che L, & Zhao H. (2024). Selenomethionine alleviates chronic heat stress-induced breast muscle injury and poor meat quality in broilers via relieving mitochondrial dysfunction and endoplasmic reticulum stress. Animal Nutrition, 16, 363-375.

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Quantifying Tissue Energy Metabolism

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For liver and muscle ATP content analysis, 12 individual fish from each genotype were sampled (two fish tissues for one sample, six replicates for each genotype) and measured according to the instructions of the assay kit (A095-1-1, Nanjing Jiancheng Bioengineering Institute, China). The results were standardized using a protein concentration kit (A045-3, Nanjing Jiancheng Bioengineering Institute, China).

Xi L., Zhai G., Liu Y., Gong Y., Lu Q., Zhang Z., Liu H., Jin J., Zhu X., Yin Z., Xie S, & Han D. (2023). Attenuated glucose uptake promotes catabolic metabolism through activated AMPK signaling and impaired insulin signaling in zebrafish. Frontiers in Nutrition, 10, 1187283.

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Quantifying Cardiac ATP Levels

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The whole heart was homogenized and adenosine triphosphate (ATP) was quantitatively determined using an ATP colorimetric assay kit (#A095-1-1, Nanjing Jiancheng Co., Nanjing, China) following the manufacturer's instructions. Briefly, the homogenates of heart tissues (200 mg in 1.8 mL of milli-Q H₂O) were heated to 100°C for 10 min and centrifuged at 1150 × g for 10 min. The amount of phosphorylating glycerol generated from the supernatants was read at 630 nm using a spectrophotometer (Ultrospec 2100, Biochrom, Cambridge, UK).

Li X., Tan W., Zheng S., Zhang J., Zhu C., Cai C., Chen H., Yang C., Kang L., Pan Z., Pyle W.G., Backx P.H., Zou Y, & Yang F.H. (2022). Cardioprotective Effects of n-3 Polyunsaturated Fatty Acids: Orchestration of mRNA Expression, Protein Phosphorylation, and Lipid Metabolism in Pressure Overload Hearts. Frontiers in Cardiovascular Medicine, 8, 788270.

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Quantification of Cardiac ATP Levels

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Fresh embryonic hearts were homogenized by a homogenizer in 4% perchloric acid and then centrifuged at 12000 rpm for 10 min. Add 1 M K₂HPO₄ into supernatant followed by another centrifugation at 12000 rpm for 10 min. The resulting supernatant was filtered by 0.22 μm membrane for high-performance liquid chromatography (HPLC) analysis. Detection conditions: UV detection wavelength: 254 nm; column: 5 μm Cosmosil C18 reversed-phase column (250 mm × 4.6 mm, i. d.); sample size: 20 μL; column temperature: room temperature; mobile phase: 50 mM potassium dihydrogen phosphate buffers (pH 6.5); flow rate: 1 mL/min. The ATP peak of samples was determined in reference to the ATP standards. The amount of ATP in samples was determined based on the regression equation from ATP standard. For the evaluation of ATP content of H9c2 cells, a commercial kit was used according to manufacturer’s instructions (A095-1-1, Nanjing Jiancheng Bioengineering Institute, Nanjing, China). ATP levels were normalized to protein content.

Yan C.Y., Ye Y., Mu H.L., Wu T., Huang W.S., Wu Y.P., Sun W.Y., Liang L., Duan W.J., Ouyang S.H., Huang R.T., Wang R., Sun X.X., Kurihara H., Li Y.F, & He R.R. (2023). Prenatal hormone stress triggers embryonic cardiac hypertrophy outcome by ubiquitin-dependent degradation of mitochondrial mitofusin 2. iScience, 27(1), 108690.

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Colorimetric Determination of Muscle ATP

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The ATP content in muscle tissues was determined via the phosphomolybdic acid colorimetric method, according to the manufacturer’s instructions (A095-1-1, Nanjing Jiancheng Bioengineering Institute, Nanjing, China).

Zhang W., You B., Qi D., Qiu L., Ripley-Gonzalez J.W., Zheng F., Fu S., Li C., Dun Y, & Liu S. (2021). Trimetazidine and exercise provide comparable improvements to high fat diet-induced muscle dysfunction through enhancement of mitochondrial quality control. Scientific Reports, 11, 19116.

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