Intracellular Oxygen Concentration Assay (ab197245)
Key features and details
- Assay type: Quantitative
- Detection method: Fluorescent
- Platform: Microplate reader
- Assay time: 1 hr 30 min
- Sample type: Adherent cells, Suspension cells
概述
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产品名称
Intracellular Oxygen Concentration Assay -
检测方法
Fluorescent -
样品类型
Adherent cells, Suspension cells -
检测类型
Quantitative -
检测时间
1h 30m -
种属反应性
与反应: Human
预测可用于: Mammals -
产品概述
Intracellular Oxygen Concentration Assay (ab197245) is an easy mix and measure 96 well fluorescence plate reader based approach for the analysis of intracellular oxygen concentration at the cell monolayer. The assay is based on the ability of oxygen to quench the excited state of the oxygen-sensitive probe. The probe is taken up via nonspecific energy dependent endocytosis and, after washing, the cells are monitored on a fluorescence plate reader (dual-read TR-F required for full oxygen quantitation). The probe phosphorescence is quenched by intracellular oxygen in a non-chemical, reversible manner allowing the measurement of average intracellular O2 levels and facilitating real-time monitoring of relative changes in cellular oxygen consumption. The probe signal increases with a reduction in intracellular oxygen and deceases with an increase in intracellular oxygen. The probe is excitable at 360-400 or 535 nm and emits at 630-680 nm. Optimal filter combinations are Ex/Em = 340/642 nm.
The flexible plate reader format, allows multiparametric or multiplex combination with a range of other reagents and it is suitable for HTP automation.
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说明
Learn more about the full range of assays to measure glycolysis, oxygen consumption, fatty acid oxidation and metabolic flux in live cells.
Or review the full metabolism assay guide for other assays for metabolites, metabolic enzymes, mitochondrial function, and oxidative stress.
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平台
Microplate reader
性能
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存放说明
Store at +4°C. Please refer to protocols. -
组件 96 tests 4 x 96 tests Intracellular O2 probe 1 vial 4 vials -
研究领域
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图片
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Excitation and Emission spectra of Intracellular O2 probe, showing normalized excitation (Ex 360-400nm; Peak 380nm) and emission (Em 630-670nm; Peak 650nm).
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HEK293T cell oxygenation. HEK293T cells were cultured in 2D and measured at ambient oxygen. Intracellular O2 levels were ~ 14%. Reducing instrument O2 to 6% caused cellular oxygenation to drop to ~2%. Assay performed using a CLARIOstart equipped with an ACU module (BMG Labtech).
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Sample Calibration Data. Intra O2 probe Lifetime profiles measured at decreasing [O2] with parallel glucose oxidase treatment to achieve 0% O2.
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Relationship between probe lifetime (τ) and applied [O2]. Applying a first order exponential fit generates a calibration function of O2% = A1 x Exp(-τ / t1). Example: O2% = 659.3 x Exp(-τ / 8.475).
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Measuring the impact of cell metabolism on iPS-derived cardiomyocyte oxygenation. During measurement, cells are treated with antimycin (ETC inhibitor) and isoproterenol (β-adrenoreceptor agonist).
数据表及文件
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SDS download
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Datasheet download
文献 (3)
ab197245 被引用在 3 文献中.
- Uribe-Alvarez C et al. Wolbachia pipientis grows in Saccharomyces cerevisiae evoking early death of the host and deregulation of mitochondrial metabolism. Microbiologyopen 8:e00675 (2019). PubMed: 29897678
- Cubillos-Zapata C et al. Obstructive Sleep Apnea Monocytes Exhibit High Levels of Vascular Endothelial Growth Factor Secretion, Augmenting Tumor Progression. Mediators Inflamm 2018:7373921 (2018). PubMed: 29997451
- Hernández-Jiménez E et al. Monocytes inhibit NK activity via TGF-ß in patients with obstructive sleep apnoea. Eur Respir J 49:N/A (2017). PubMed: 28619958