Carboxy-H2DCFDA (indicateur de stress oxydatif général)

Citations et références (95)

Invitrogen™

Carboxy-H2DCFDA (indicateur de stress oxydatif général)

Référence	Quantité
C400	25 mg

Référence C400

Prix (EUR)

435,65

Precio exclusivo en nuestra web

474,00

Économisez 38,35 (8%)

Quantité:

25 mg

Prix (EUR)

435,65

Precio exclusivo en nuestra web

474,00

Économisez 38,35 (8%)

Le diacétate de 6-carboxy-2’,7’-dichlorodihydrofluorescéine est chimiquement réduit, sous forme acétylée de fluorescéine utilisée comme indicateur pour les dérivés réactifs de l’oxygène (DRO) dans les cellules. Cette molécule non fluorescente est facilement convertie en fluorescence verte lorsque les groupes d’acétate sont éliminés par des estérases intracellulaires et que l’oxydation (par l’activité des DRO) se produit à l’intérieur de la cellule. Nous proposons l’analogue carboxylique H2DCFDA (carboxy-H2DCFDA , C400), qui a deux charges négatives à pH physiologique, et son di(acétoxyméthyl ester) (C2938). Lors du clivage des groupes acétate et ester par des estérases intracellulaires et par oxydation, les deux analogues forment de la carboxydichlorofluorescéine (C368), avec des charges négatives supplémentaires qui empêchent sa fuite hors de la cellule.

Spécifications de l’indicateur DRO :
• Ex / Em : ∼ 492 – 495/517 – 527 nm
• Le produit est sensible à l’air et doit être stocké sous argon sec ou azote
• Le produit peut être dissous dans DMSO, DMF, ou éthanol à utiliser
• L’indicateur est perméant aux cellules (les protocoles de chargement cellulaire sont disponibles dans la littérature)
• La fluorescence peut être surveillée à l’aide d’un cytomètre en flux, d’un fluoromètre, d’un lecteur de microplaques ou d’un microscope à fluorescence, utilisation de sources d’excitation et de filtres adaptés à la fluorescéine

En savoir plus sur les indicateurs DRO
Nous proposons un assortiment de produits Molecular Probes™ pour la génération d’espèces réactives d’oxygène (DRO), y compris l’oxygène singulet, le superoxyde, le radical hydroxyle et divers peroxyde et hydroperoxydes, ainsi que pour leur détection fluorométrique en solution. Consultez Génération et détection des espèces réactives à l’oxygène : Section 18.2 dans le manuel sur les sondes moléculaires™ si vous souhaitez plus d’informations sur ces produits.

Réservé à la recherche. Non utilisé à des fins thérapeutiques ou diagnostiques humaines ou animales.

Usage exclusivement réservé à la recherche. Ne pas utiliser pour des procédures de diagnostic.

Spécifications

Quantité25 mg

Type de produitROS Indicator

Unit SizeEach

Foire aux questions (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.

Citations et références (95)

Citations et références

Abstract

N-acetylcysteine augments the cellular redox changes and cytotoxic activity of internalized mycobacterium bovis in human bladder cancer cells.

Authors:Pook SH, Esuvaranathan K, Mahendran R

Journal:J Urol

PubMed ID:12131368

PURPOSE: We determined whether changes in cellular reactive oxygen species correlated with mycobacteria internalization and bladder cancer cell death. MATERIALS AND METHODS: Reactive oxygen species and thiols in RT112 and MGH bladder cancer cells were determined using the fluorescence probes 5-(and 6)-carboxy-2', 7' dichlorodihydrofluorescein diacetate and monobromobimane. Superoxide and nitrite ... More

The sulphonylurea glibenclamide inhibits multidrug resistance protein (MRP1) activity in human lung cancer cells.

Authors:Payen L, Delugin L, Courtois A, Trinquart Y, Guillouzo A, Fardel O

Journal:Br J Pharmacol

PubMed ID:11159731

'1. Glibenclamide, a sulphonylurea widely used for the treatment of non-insulin-dependent diabetes mellitus, has been shown to inhibit the activities of various ATP-binding cassette (ABC) transporters. In the present study, its effects towards multidrug resistance protein 1 (MRP1), an ABC efflux pump conferring multidrug resistance and handling organic anions, were ... More

Characterization of Vta1p, a class E Vps protein in Saccharomyces cerevisiae.

Authors:Shiflett SL, Ward DM, Huynh D, Vaughn MB, Simmons JC, Kaplan J

Journal:J Biol Chem

PubMed ID:14701806

'We identified VTA1 in a screen for mutations that result in altered vacuole morphology. Deletion of VTA1 resulted in delayed trafficking of the lipophilic dye FM4-64 to the vacuole and altered vacuolar morphology when cells were exposed to the dye 5-(and 6)-carboxy-2'',7''-dichlorofluorescein diacetate (CDCFDA). Deletion of class E vacuolar protein ... More

Mitochondrial localization of reactive oxygen species by dihydrofluorescein probes.

Authors:Diaz G, Liu S, Isola R, Diana A, Falchi AM

Journal:Histochem Cell Biol

PubMed ID:14574587

'Mitochondria are the main source of reactive oxygen species (ROS). The aim of this work was to verify the ROS generation in situ in HeLa cells exposed to prooxidants and antioxidants (menadione, tert-butyl hydroperoxide, antimycin A, vitamin E, N-acetyl-L-cysteine, and butylated hydroxytoluene) using the ROS-sensitive probes 6-carboxy-2'',7''-dichlorodihydrofluorescein diacetate di-acetomethyl ester ... More

The use of fluorescent probes to assess oxidative processes in isolated-perfused rat heart tissue.

Authors:Kehrer JP, Paraidathathu T

Journal:Free Radic Res Commun

PubMed ID:1505782

'The formation of reactive oxygen species (ROS) in intact heart tissue has been assessed by direct ESR measurements, and indirectly by the formation of characteristic tissue products and the protective effects of various antioxidants. The development of lipid soluble esters of compounds which can be trapped intracellularly after hydrolysis, and ... More

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