CM-H2DCFDA (allgemeiner Oxidativ-Stress-Indikator)

Zitierungen und Referenzen (115)

Invitrogen™

CM-H2DCFDA (allgemeiner Oxidativ-Stress-Indikator)

CM-H 2DCFDA ist ein Chlormethylderivat von H 2DCFDA und ein nützlicher Indikator für reaktive Sauerstoffspezies (ROS) in Zellen. Dieser IndikatorWeitere Informationen

Katalognummer	Menge
C6827	20 x 50 μg

Katalognummer C6827

Preis (EUR)

644,65

Exklusiv online

686,00

Ersparnis 41,35 (6%)

Zum Warenkorb hinzufügen

Menge:

20 x 50 μg

Preis (EUR)

644,65

Exklusiv online

686,00

Ersparnis 41,35 (6%)

Zum Warenkorb hinzufügen

CM-H₂DCFDA ist ein Chlormethylderivat von H₂DCFDA und ein nützlicher Indikator für reaktive Sauerstoffspezies (ROS) in Zellen. Dieser Indikator zeigt eine weitaus bessere Retention in lebenden Zellen als H₂DCFDA. CM-H₂DCFDA diffundiert passiv in die Zellen, wo seine Acetatgruppen durch intrazelluläre Esterasen gespalten werden und seine thiol-reaktive Chlormethylgruppe mit intrazellulärem Glutathion und anderen Thiolen reagiert. Das durch anschließende Oxidation entstehende fluoreszierende Addukt wird in der Zelle eingeschlossen und eignet sich deshalb für Langzeitstudien.

Spezifikationen des ROS-Indikators:

• Ex/Em: ∼492 – 495/517 – 527 nm
• Produkt ist luftempfindlich und sollte unter trockenem Argon oder Stickstoff gelagert werden
• Produkt kann zur Verwendung in DMSO, DMF oder Ethanol gelöst werden
• Membrangängiger Indikator (Zellladeprotokolle sind in den Literaturhinweisen erhältlich)
• Die Fluoreszenz kann durch Durchflusszytometer, Fluorometer, Mikrotiterplatten-Lesegeräte oder Fluoreszenzmikroskope mit Anregungsquellen und Filtern für Fluorescein überwacht werden

Weitere ROS-Indikatoren
Wir bieten ein Sortiment an Molecular Probes™ Produkten für die Herstellung reaktiver Sauerstoffspezies (ROS) an, einschließlich Singulett-Sauerstoff, Superoxid, Hydroxyl-Radikal und verschiedenen Peroxiden und Hydroperoxiden sowie für deren fluorometrischen Nachweis in Lösung. Weitere Informationen zu diesen Produkten finden Sie im Abschnitt Herstellung und Nachweis reaktiver Sauerstoffspezies – Abschnitt 18.2 im Molecular Probes™ Handbuch.

Nur für Forschungszwecke. Nicht zur Verwendung bei diagnostischen Verfahren.

Specifications

Menge20 x 50 μg

ProdukttypROS Indicator

Unit SizeEach

Häufig gestellte Fragen (FAQ)

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.

Zitierungen und Referenzen (115)

Zitierungen und Referenzen

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

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