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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.
|组件||96 tests||4 x 96 tests|
|Intracellular O2 probe||1 vial||1 x 4 vials|
Our Abpromise guarantee covers the use of ab197245 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Cellular activity||Use at an assay dependent concentration.|
Excitation and Emission spectra of Intracellular O2 probe, showing normalized excitation (Ex 360-400nm; Peak 380nm) and emission (Em 630-670nm; Peak 650nm).
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).
Sample Calibration Data. Intra O2 probe Lifetime profiles measured at decreasing [O2] with parallel glucose oxidase treatment to achieve 0% O2.
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).
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).
ab197245 has not yet been referenced specifically in any publications.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"