CellROX™ Tiefrotes Reagenz für den Nachweis von oxidativem Stress

Zitierungen und Referenzen (52)

Invitrogen™

CellROX™ Tiefrotes Reagenz für den Nachweis von oxidativem Stress

Das CellROX™ Tiefrot Reagenz ist ein neuartiger fluorogener Farbstoff zur Messung des zellulären oxidativen Stress sowohl lebender als auch fixierterWeitere Informationen

Katalognummer	Menge
C10422	250 μl

Katalognummer C10422

Preis (EUR)

633,65

Exklusiv online

674,00

Ersparnis 40,35 (6%)

Menge:

250 μl

Preis (EUR)

633,65

Exklusiv online

674,00

Ersparnis 40,35 (6%)

Das CellROX™ Tiefrot Reagenz ist ein neuartiger fluorogener Farbstoff zur Messung des zellulären oxidativen Stress sowohl lebender als auch fixierter Zellen, mit einem Absorptions-/Emissionsmaximum bei ∼644/665 nm. Der zellgängige Farbstoff fluoresziert in reduziertem Zustand nicht und zeigt nach Oxidation durch die reaktive Sauerstoffspezies (ROS) eine helle Fluoreszenz.

Merkmale von CellROX™ tiefrotem Reagenz:

• Optimierte Sonde für den Nachweis von oxidativem Stress in Zellen
• Einfaches Protokoll, das mit anderen Lebendzell-Farbstoffen und GFP verwendet werden kann
• Kompatibel mit der Fluoreszenz-Bildgebung lebender Zellen und mit Formaldehyd-basierten Fixierverfahren

Substrateigenschaften
Das CellROX™ tiefrote Reagenz ist ein neuer, membrangängiger Farbstoff mit Absorptions-/Emissionsmaxima von ∼644/665 nm. CellROX™ tiefrotes Reagenz ist in reduziertem Zustand nicht fluoreszierend und zeigt bei Oxidation durch reaktive Sauerstoffspezies eine Fluoreszenz mit einem Emissionsmaximum von ∼665 nm, die durch Fluoreszenz-Bildgebung, High-Content-Bildgebung, Fluoreszenz-Platten-Lesegeräte und Durchflusszytometrie messbar ist.

Nachweis von oxidativem Stress mit CellROX™ tiefrotem Reagenz
Oxidativer Stress entsteht durch ein Ungleichgewicht zwischen der Produktion reaktiver Sauerstoffspezies (ROS) und der Fähigkeit der Zellen, diese zu neutralisieren. ROS spielen eine wichtige Rolle beim Fortschreiten verschiedener Erkrankungen, zu denen Entzündungen, Atherosklerose, Alterungsprozesse und altersbedingte degenerative Erkrankungen gehören. CellROX™ tiefrotes Reagenz kann den oxidativen Stress in Zellen nachweisen, indem es mit ROS reagiert und so hell fluoreszierend wird.

Einfaches, robustes Protokoll
Das Protokoll für die Verwendung dieses Reagenz ist einfach und das helle, tiefrote Fluoreszenzsignal ist mit anderen Lebendzell-Farbstoffen und GFP kompatibel. Dadurch eignet es sich für Multiplex-Fluoreszenz-Assays zur Messung einer Vielzahl von zellulären Phänomenen einschließlich Parametern in Bezug auf Zytotoxizität und Zelltod (Abbildung 4 unten). Außerdem bleibt das Signal von CellROX™ Tiefrot nach der Formaldehyd-Fixierung, im Gegensatz zu vielen anderen ROS-Sensoren wie H2DCFDA, erhalten. Dies ermöglicht Flexibilität mit dem Assay und verbesserte Workflows im Vergleich zu denen, die auf klassischen Farbstoffen für den ROS-Nachweis basieren.

Nur für Forschungszwecke. Nicht zur Verwendung bei diagnostischen Verfahren.

Specifications

FarbeTiefrot

Konzentration2.5 mM stabilized solution in DMSO

Zur Verwendung mit (Geräte)Imaging, HCS, Cytometer

FormatFlüssig

Menge250 μl

NachweisverfahrenLive Cell Imaging

Excitation/Emission644/665 nm.

IndicatorOxidative stress

ProduktlinieCellROX

Unit SizeEach

Inhalt und Lagerung

Store at ≤–20°C. Protect from light and desiccate.

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 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.

What dyes can I use to detect reactive oxygen species (ROS) in my bacteria?

Many dyes that are used on mammalian cells have also been shown to be useful in bacterial cells. For example, CellROX Deep Red Reagent has been shown to work in B. subtilis (see Reference: http://www-brs.ub.ruhr-uni-bochum.de/netahtml/HSS/Diss/RaatschenNadja/diss.pdf). If you are interested in a particular dye, but are not sure if it will work on your bacteria, literature searches are the best way to check to see if it has been tested. If not, then it may be worth testing yourself.

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

I am trying to label my cells with a reactive oxygen species (ROS) indicator dye, but I am not seeing a significant difference in signal. What could be happening?

First, make sure you have both a negative (untreated) and positive (ROS-induced) sample to compare. A good positive control can be the use of 100 µM menadione for one hour or 50 µM nefazodone for 24 hours. H2O2 can also be used, though it does not work well for CellROX dyes. Some dyes, such as H2DCFDA, require esterase cleavage, so don't incubate in the presence of serum (which contains esterases that can prematurely cleave the dye). If your positive control does not show significant change compared to the negative control, try increasing the concentration and label time for the dye. Our manuals give starting recommendations. Be sure to image your live cells as soon as possible. Only two dyes (CellROX Green and CellROX Deep Red) are retained with formaldehyde fixation. Finally, make sure you are using filters and instrument settings to match the excitation and emission spectra of the dye.

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

Zitierungen und Referenzen (52)

Zitierungen und Referenzen

Abstract

The airway epithelium nucleotide-binding domain and leucine-rich repeat protein 3 inflammasome is activated by urban particulate matter.

Authors:Hirota JA, Hirota SA, Warner SM, Stefanowicz D, Shaheen F, Beck PL, Macdonald JA, Hackett TL, Sin DD, Van Eeden S, Knight DA,

Journal:J Allergy Clin Immunol

PubMed ID:22227418

The airway epithelium is the first line of defense against inhaled insults and therefore must be capable of coordinating appropriate inflammatory and immune responses. We sought to test the hypothesis that the nucleotide-binding domain and leucine-rich repeat protein 3 (NLRP3) inflammasome, an intracellular danger-sensing complex, plays a critical role in ... More

Combination Small Molecule MEK and PI3K Inhibition Enhances Uveal Melanoma Cell Death in a Mutant GNAQ- and GNA11-Dependent Manner.

Authors:Khalili JS, Yu X, Wang J, Hayes BC, Davies MA, Lizee G, Esmaeli B, Woodman SE,

Journal:Clin Cancer Res

PubMed ID:22733540

Activating Q209L/P mutations in GNAQ or GNA11 (GNAQ/11) are present in approximately 80% of uveal melanomas. Mutant GNAQ/11 are not currently therapeutically targetable. Inhibiting key down-stream effectors of GNAQ/11 represents a rational therapeutic approach for uveal melanomas that harbor these mutations. The mitogen-activated protein/extracellular signal-regulated kinase/mitogen-activated protein kinase (MEK/MAPK) and ... More

Mutant BRAF induces DNA strand breaks, activates DNA damage response pathway, and up-regulates glucose transporter-1 in nontransformed epithelial cells.

Authors:Sheu JJ, Guan B, Tsai FJ, Hsiao EY, Chen CM, Seruca R, Wang TL, Shih IeM,

Journal:Am J Pathol

PubMed ID:22227015

'Although the oncogenic functions of activating BRAF mutations have been clearly demonstrated in human cancer, their roles in nontransformed epithelial cells remain largely unclear. Investigating the cellular response to the expression of mutant BRAF in nontransformed epithelial cells is fundamental to the understanding of the roles of BRAF in cancer ... More

Mechanisms of programmed cell death signaling in hair cells and support cells post-electrode insertion trauma.

Authors:Eshraghi AA, Lang DM, Roell J, Van De Water TR, Garnham C, Rodrigues H, Guardiola M, Gupta C, Mittal J,

Journal:

PubMed ID:25761716

'Programmed cell death (PCD) initially starts in the support cells (SCs) after electrode insertion trauma (EIT), followed by PCD in hair cells (HCs). Activation of caspase-3 was observed only in SCs. Protecting both SCs and HCs with selective otoprotective drugs at an early stage post implantation may help to preserve ... More

OPA1 Mutation and Late-Onset Cardiomyopathy: Mitochondrial Dysfunction and mtDNA Instability.

Authors:Chen L, Liu T, Tran A, Lu X, Tomilov AA, Davies V, Cortopassi G, Chiamvimonvat N, Bers DM, Votruba M, Knowlton AA,

Journal:J Am Heart Assoc

PubMed ID:23316298

'Mitochondrial fusion protein mutations are a cause of inherited neuropathies such as Charcot-Marie-Tooth disease and dominant optic atrophy. Previously we reported that the fusion protein optic atrophy 1 (OPA1) is decreased in heart failure. We investigated cardiac function, mitochondrial function, and mtDNA stability in a mouse model of the disease ... More

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