CM-H2DCFDA (indicador de estrés oxidativo general)

Citas y referencias (115)

Invitrogen™

CM-H2DCFDA (indicador de estrés oxidativo general)

CM-H2DCFDA es un derivado clorometílico de H2DCFDA, que resulta útil como indicador de especies reactivas de oxígeno (ROS) en células.Más información

Número de catálogo	Cantidad
C6827	20 × 50 μg

Número de catálogo C6827

Precio (MXN)

Cantidad:

20 × 50 μg

CM-H₂DCFDA es un derivado clorometílico de H₂DCFDA, que resulta útil como indicador de especies reactivas de oxígeno (ROS) en células. Este indicador muestra un mayor nivel de retención en células vivas que H₂DCFDA. CM-H₂DCFDA se expande pasivamente en las células, en las que las esterasas intracelulares escinden los grupos de acetato. Su grupo de clorometilo reactivo al tiol reacciona con el glutatión y otros tioles. La oxidación posterior produce un aducto fluorescente que queda atrapado dentro de la célula, por lo que facilita los estudios a largo plazo.

Especificaciones del indicador de ROS:

• Ex/Em: ∼492–495/517–527 nm
• Este producto es sensible al aire y se debe almacenar bajo nitrógeno o argón seco
• Este producto se puede disolver en DMSO, DMF o etanol para su uso
• El indicador es capaz de penetrar en la célula (en la documentación, se incluyen protocolos de carga de células)
• La fluorescencia puede monitorizarse mediante un citómetro de flujo, un fluorímetro, un lector de microplacas o un microscopio de fluorescencia a través de fuentes de excitación y filtros adecuados para la fluoresceína

Encuentre más indicadores de ROS
Ofrecemos un surtido de productos de Molecular Probes™ para la generación de especies reactivas de oxígeno (ROS), incluido el oxígeno singulete, el superóxido, el radical hidroxilo y diversos peróxidos e hidroperóxidos, así como para su detección fluorimétrica en solución. Consulte el apartado de generación y detección de especies reactivas de oxígeno (sección 18.2) en el manual de Molecular Probes™ para obtener más información sobre estos productos.

Para uso exclusivo en investigación. No apto para uso en procedimientos diagnósticos.

Especificaciones

Cantidad20 × 50 μg

Tipo de productoROS Indicator

Unit SizeEach

Preguntas frecuentes

I want to assay cells for reactive oxygen species using carboxy-H2DCFDA, but I want to do so with a plate reader instead of microscope. Will it work?

It has been done. The problem is that plate readers are less sensitive than microscopes, with far less signal-to-background difference. It is worth trying, but first optimize concentrations and loading times with control cells, use a plate with little to no autofluorescence, and possibly optimize the gain setting in order to get the best signal possible. But don't expect the same sensitivity, even with optimization.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

I have GFP-transfected cells and need to label for reactive oxygen species. Can I use H2DCFDA?

This is not recommended as the two dyes overlap in the emission wavelength. There are other ROS reagents available in different wavelengths, such as CellROX Deep Red, which emits in the far-red range (665 nm), or dihydroethidium, which is emits in the visible red range (620 nm).

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

I labeled my cell with CM-H2DCFDA for reactive oxygen detection, but upon illuminating the cell there is a significant increase in fluorescence in the control cells. Why?

If the cell is overloaded with dye, the high intracellular concentration of the dye may lead to dye-dye quenching. Upon illumination, photobleaching will occur, which will reduce the dye-dye quenching and actually increase the fluorescence (for a while, but then it will start decreasing). To solve the problem, reduce the concentration and incubation time, and try a range of incubation times and concentrations.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

I need a formaldehyde-fixable reactive oxygen species detection assay. Is H2 DCFDA fixable?

H₂DCFDA and similar derivatives are not fixable. The same goes for dihydroethidium and dihydrorhodamine. However, CellROX Deep Red and CellROX Green are retained for a limited time upon fixation with formaldehyde. CellROX Green may be retained upon subsequent Triton X-100 permeabilization. Avoid the use of any acetone or alcohol-based fixatives or fixatives that include alcohol, such as formalin.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

Why don't I see a significant change in signal for my live-cell fluorescent indicator dye?

Regardless of the type of live-cell indicator dye (e.g., calcium indicators, pH indicator, metal ion indicators), make sure there is no serum during the loading step, which can prematurely cleave dyes with AM esters and bind dyes non-specifically. Always optimize the dye concentration and staining time with a positive control before you run your test samples, to give the best signal-to-background. Always run a positive control with a buffer containing free ions of known concentration and an ionophore to open pores to those ions (for instance, for calcium indicators like Fluo-4 AM, this would include a buffer with added calcium combined with calcimycin, or for pH indicators, buffers of different pHs combined with nigericin). Reactive oxygen indicators, such as CellROX Green or H2DCFDA would require a cellular reactive oxygen species (ROS) stimulant as a positive control, such as menadione. Finally, make sure your imaging system has a sensitive detector. Plate readers, for instance, have much lower detector efficiency over background, compared to microscopy or flow cytometry.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

Citations & References (115)

Citations & References

Abstract

Authors:

Journal:

PubMed ID:18258751

Large-scale chemical dissection of mitochondrial function.

Authors:Wagner BK,Kitami T,Gilbert TJ,Peck D,Ramanathan A,Schreiber SL,Golub TR,Mootha VK

Journal:Nature biotechnology

PubMed ID:18297058

Mitochondrial oxidative phosphorylation (OXPHOS) is central to physiology and disease pathogenesis. To systematically investigate its activity and regulation, we performed a wide range of assays of OXPHOS physiology and nuclear and mitochondrial gene expression across 2490 chemical perturbations in muscle cells. Through mining of the resulting compendium, we discovered that: ... More

Reactive oxygen species production via NADPH oxidase mediates TGF-beta-induced cytoskeletal alterations in endothelial cells.

Authors:Hu T, Ramachandrarao SP, Siva S, Valancius C, Zhu Y, Mahadev K, Toh I, Goldstein BJ, Woolkalis M, Sharma K

Journal:Am J Physiol Renal Physiol

PubMed ID:16159901

'Cytoskeletal alterations in endothelial cells have been linked to nitric oxide generation and cell-cell interactions. Transforming growth factor (TGF)-beta has been described to affect cytoskeletal rearrangement in numerous cell types; however, the underlying pathway is unclear. In the present study, we found that human umbilical vein endothelial cells (HUVEC) have ... More

The yeast prion Ure2p native-like assemblies are toxic to mammalian cells regardless of their aggregation state.

Authors:Pieri L, Bucciantini M, Nosi D, Formigli L, Savistchenko J, Melki R, Stefani M

Journal:J Biol Chem

PubMed ID:16571726

'The yeast prion Ure2p assembles in vitro into oligomers and fibrils retaining the alpha-helix content and binding properties of the soluble protein. Here we show that the different forms of Ure2p native-like assemblies (dimers, oligomers, and fibrils) are similarly toxic to murine H-END cells when added to the culture medium. ... More

Role of extracellular signal-regulated protein kinase in neuronal cell death induced by glutathione depletion in neuron/glia mesencephalic cultures.

Authors:de Bernardo S, Canals S, Casarejos MJ, Solano RM, Menendez J, Mena MA

Journal:J Neurochem

PubMed ID:15485497

'To date, glutathione (GSH) depletion is the earliest biochemical alteration shown in brains of Parkinson''s disease patients, but the role of GSH in dopamine cell survival is debated. In this study we show that GSH depletion, produced with GSH synthesis inhibitor, L-buthionine-(S,R)-sulfoximine (BSO), induces selectively neuronal cell death in neuron/glia, ... More

View all CM-H2DCFDA (indicador de estrés oxidativo general) citations