(F) ROS levels were assessed using ROS test kit. Popular MitoSOX Red MitoSOX Red Antibodies. Optical Data for Mito-HE Plan Your Experiments Use our spectra viewer to interactively plan your experiments, assessing multiplexing options. ZERO BIAS - scores, article reviews, HUVEC (green) and the J-aggregates (red) were detected at 530 nm and 590 nm emission, respectively. Therefore, we stained MitoSOX Excitation Emission Maximum, supplied by Thermo Fisher, used in various techniques. MitoSOX superoxide indicators are novel fluorogenic dyes specifically targeted to mitochondria in live cells. It is recommended to prepare stock solutions in DMSO. 750 nm and 810 nm NIR interferes with the excitation/emission spectra for these fluorescent indicators and thus precluded us from testing these wavelengths for parallel effects. It is a new type of fluorescent probe specifically targeting live cell line sites. A collection of spectra representing the microscope Leica Thunder Imager 3D cell culture BIU on FPbase. Thermo Fisher mitosox red Mitosox Red, supplied by Thermo Fisher, used in various techniques. The maximum excitation/emission wavelength is 651/670 nm. Mitochondrial Respiration The ROS probe channels were recorded under 514 nm excitation by collecting emission at 570690 nm (MitoSox red) or 530650 nm (SOSG and APF). CellEvent Caspase-3/7 Red Detection Reagent: absorption/emission maxima of 590/610 nm (traditional Texas red filter set) How to use CellEvent Caspase-3/7 detection reagents are 2.6. (CE) MDA, SOD, and GSH levels were measured by corresponding test kits. MitoSOX red superoxide indicator has maximum excitation and emission at 510 and 580 nm, respectively. 1) 50 ug of lyophilized MitoTracker Deep Red was diluted in 100 ul of DMSO to produce 0.92mM stock (kept in -20C) 2) 100 nM and 200 nM working solution was prepared, diluted in Opti Company / SKU Applications Marker / Reactivity Host / Isotype / Clone Size / Price Excitation/emission maxima ~ 510/580 nm. Mitochondrial aconitase, the tricarboxylic acid cycle (TCA) enzyme, was identified as the main target of excitotoxic O 2 in cortical cultures [ 17 ], which inactivation closely correlated with subsequent neuronal death [ 20 ]. Scale bar, 50 m. Oxidation of the MitoSOX reagent by mitochondrial superoxide produces bright The ideal staining method is to stain 30 minutes at 37 degree celcius before fixation with 4% paraformaldehyde at room temperature. It has membrane permeability and produces strong Red The excitation/emission fluorescence for calcein-AM is 494/517 nm. Preparation of Reagents MitoSOX reagent solution (2.5 mM in DMSO): 50 g MitoSOX dissolved in 26 l DMSO (store at 20C). ZERO BIAS - scores, MitoSOX Red reagent is a novel fluorogenic dye specifically targeted to mitochondria in live cells. Oxidation of MitoSOX Red reagent by superoxide produces red fluorescence. Readily oxidized by superoxide but not by other ROS- or RNS-generating systems Absorption/emission maxima: ~510/580 nm Fluorescence intensities from seven randomly selected fields in each dish were measured using the Oxidation of the MitoSOX reagent by mitochondrial superoxide produces bright green or red fluorescence. Once entering mitochondria, it MitoSOX Red Excitation and Emission Spectra. Remove from water bath and add 0.5 ml of Bioz Stars score: 99/100, based on 1 PubMed citations. Since MitoTracker Red and MitoSOX have very similar excitation and emission wavelengths, these two probes cannot be used together. Mitochondrial superoxide formation was detected by incubating cells in the dark with 5 m MitoSOX Red dye (excitation/emission at = 510 nm/580 nm) for 30 min. The integrated density of MitoSOX staining was achieved using NIH ImageJ software analysis, and the results were represented as arbitrary fluorescence units. Bioz Stars score: 99/100, based on 1 PubMed citations. MitoSOX Red exhibits fluorescence after oxidation by superoxide anion (excitation 510 nm, emission 580 nm) . 4.3. ZERO BIAS - scores, This incubation time can be increased to 40 min depending on experiments. Detection of MitoSOX Red indicator by flow cytometry typically uses the FL2 emission channel (585/42 nm). Incubate all 7 tubes in a shaking, 37C water bath for 20 min. (1) reported that if MitoSOX Red dye is excited at 396 nm instead of 510 nm, it can be a more selective marker for mitochondrial superoxide. MitoSOX Red reagent is readily oxidized by tation/emission maxima of approximately 510/580 nm. The Cy5 HYQ combination employs a longpass dichromatic mirror having a cut-on wavelength of 660 nanometers, which is 10 nanometers higher than the excitation bandpass cut-off. Stained cells were analyzed using an Attune NxT acoustic focusing flow cytometer (Thermo Fisher Scientific), and flow cytometry data were analyzed using FlowJo software (v10, Tree Star). MitoSOX Red reagent is oxidized by superoxide and exhibits red fluorescence. It has membrane permeability and can bind mitochondria quickly and selectively. The MitoSOX Red Mitochondrial Superoxide Indicator (MitoSOX) is a novel fluorescent probe specifically targeted to live cell lines. However, it remains unclear how the increase evoked by NMDAR of [Ca 2+] drive O 2 formation in mitochondria. MitoSOX superoxide indicators are novel fluorogenic dyes specifically targeted to mitochondria in live cells. The MitoSOX Red mitochondrial superoxide indicator was detected using a confocal microscope at an excitation/emission maxima of 510/580 nm. For Research Use Only. Not for use in diagnostic procedures. Store in freezer (-5 to -30C) and protect from light. Convenient, on-site access to the products you need. Learn more. Detection of superoxide in live cells using MitoSOX Red superoxide indicator. There are no frequently asked questions yet. The production of superoxide by mitochondria can be visualized in fluorescence microscopy using the MitoSOX Red reagent. MitoSOX Red reagent permeates live cells where it selectively targets mitochondria. They are widely used to label peptides, proteins, and oligonucleotides, among others. MitoSOX TM Red excitation was performed at 488 nm, and emission was collected at 580 nm. MitoSox Red indicator: absorption/emission maxima of 396/610 nm* (custom filter set recommended) *Robinson et al. Excitation/Emission: 488/510 nm (green), 396/610 nm (red) Indicator: MitoSox Red indicator: absorption/emission maxima of 396/610 nm* (custom filter set recommended) *Robinson et al. DCF and MitoSOX Red fluorescence levels were measured at an excitation wavelength of 488 nm and an emission wavelength of 515540 nm. G I are intensity View the excitation and emission spectra for our fluorescent dye range and other commonly used dyes. Fluorescence intensity was calculated as a percentage by gating upon the positive and negative controls. MMP is evaluated as the red-to-green fluorescence intensity ratio. The superoxide anion reactive oxygen species (ROS) is generated as a by-product of mitochondrial oxidative phosphorylation. A wide 60-nanometer excitation band encompassing orange and red wavelengths covers the spectral range of 590 to 650 nanometers. Bioz Stars score: 93/100, based on 1 PubMed citations. MitoSOX Green and MitoSOX Red superoxide indicators are novel fluorogenic dyes specifically targeted to mitochondria in live cells. MitoSOX Red reagent is readily oxidized by tation/emission maxima of approximately 510/580 nm. (A) Mitochondrial superoxide was detected and quantified through immunofluorescence by using MitoSox Red staining. Lipid Peroxidation and Superoxide Dismutase Measurements Detection of MitoSOX Red indicator by flow cytometry typically uses the FL2 emission channel (585/42 nm). A common application of MitoSOX Red indicator is the correlation of superoxide generation with the accumulation of oxidative damage products such as lipid peroxides and modified DNA bases such as 8-oxodG. Generation of superoxide, particularly as a result of (1) reported that if MitoSOX Red dye is excited at 396 nm Cy5 maleimide is a fluorescent dye and various cyanidin 5 (Cy5) dyes have been used to label biomolecules for fluorescent imaging and other fluorescence-based biochemical analyses. To assess MitoSOX Red reagent is oxidized by superoxide and exhibits red fluorescence. Thermo Fisher mitosox red Mitosox Red, supplied by Thermo Fisher, used in various techniques. Oxidative Damage Applications A common application of MitoSOX Red The results are expressed as the real-time photon emission curve (CL response) and the integrated CL response (the area under the curve, representing the total counts of photon emission over the 60 min of incubation time). Data were analyzed by Image J software (see Leanza et al., 2017) and values at each time were expressed as percent of the value before caffeine administration. 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