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Réactifs Amplex™ Red et Amplex™ UltraRed
Citations et références (228)
Invitrogen™

Réactifs Amplex™ Red et Amplex™ UltraRed

Le réactif Amplex Red, une sonde hautement sensible et stable pour H2O2, est notre meilleur substrat fluorogénique pour la peroxydase.Afficher plus
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RéférenceQuantitéType de colorant
A122225mgRéactif Amplex Red
A2217710 x 10mgRéactif Amplex Red
A360065 x 1 mgRéactif UltraRed Amplex
Référence A12222
Prix (EUR)
354,00
Each
Quantité:
5mg
Type de colorant:
Réactif Amplex Red
Prix (EUR)
354,00
Each
Le réactif Amplex Red, une sonde hautement sensible et stable pour H2O2, est notre meilleur substrat fluorogénique pour la peroxydase. En présence de péroxydase de raifort (HRP), le réactif Amplex Red réagit en stœchiométrie : avec H2O2 pour créer une résorufine hautement fluorescente Étant donné que H2O2 est produit dans de nombreuses réactions enzymatiques différentes, le réactif Amplex Red peut être utilisé pour détecter l'activité de plusieurs enzymes différentes.
Usage exclusivement réservé à la recherche. Ne pas utiliser pour des procédures de diagnostic.
Spécifications
Méthode de détectionFluorescence
Type de colorantRéactif Amplex Red
FormeSolide
Quantité5mg
Conditions d’expéditionTempérature ambiante, Température ambiante
SubstratSubstrat HRP (péroxydase de raifort)
Propriétés du substratSubstrat chimique
Application validéeAnalyse cellulaire
À utiliser avec (application)Test de la peroxydase d’hydrogène
À utiliser avec (équipement)Fluorimètre, Lecteur de microplaques
Gamme de produitsAmplex
Type de produitPeroxyde d’hydrogène rouge
Unit SizeEach
Contenu et stockage
Conserver au congélateur (-5 à -30°C).

Foire aux questions (FAQ)

I'm using an Amplex Red kit, the reagent changes color to pink almost immediately in my own Krebs-Ringer buffer but not in HBSS. Why is this?

The components of Krebs-Ringer buffer (salts) should not cause oxidation of the Amplex reagent (which, in the presence of peroxidase and H2O2 oxidizes to resorufin, which is pink in color and fluorescent). Try water alone (the water used to make the Krebs-Ringer buffer). Since Hank's Buffered Saline Solution is typically purchased rather than made in the lab, it likely would not have the same contaminant. Another option is to degas the buffer prior to use to removed dissolved oxygen radicals.

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

Is it possible to use the Amplex Red Reagent (Cat. No. A12222) with a colorimetric microplate reader and measure the optical density (OD)? What would be the ideal wavelength for OD measurements?

Yes it can be used for a colorimetric assay. In general, absorbance is less sensitive than fluorescence so the sensitivity of the colorimetric assay may be lower. You may need to empirically determine the detection limit and linear dynamic range for the colorimetric assay. Resorufin, the fluorescent product of Amplex Red, has an absorbance peak at 570 nm.

Find additional tips, troubleshooting help, and resources within our Cell Viability, Proliferation, Cryopreservation, and Apoptosis Support Center.

I have ordered and tested your Amplex Red Reagent (Cat. Nos. A12222, A22177), but it is not reacting correctly. What could be the cause?

Either the horseradish peroxidase or other enzymes in the assay may not be active, the H2O2 may have degraded, or the sample could have some component that is consuming H2O2 or inactivating the enzymes required for the assay. We recommend that you check when the reagent was purchased and how it was stored prior to use; the reagent should not be used past the warranty period.

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

Do you recommend using Amplex Red Reagent (Cat. Nos. A12222, A22177) in live cells?

Amplex Red is cell impermeant, but also its conversion to resorufin requires direct contact with the reaction of peroxidase with H2O2 (in solution, at the surface, or within cells). Within cells, peroxidases and H2O2 may not be sufficiently localized or active to provide enough signal.

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

Can I use Amplex Red Reagent (Cat. Nos. A12222, A22177) in the presence of catalases?

Unless you are specifically analyzing catalase activity, as with the Amplex Red Catalase Assay Kit (Cat. No. A22180), the assay should be devoid of catalases. Catalases compete with the horseradish peroxidase for H2O2; the conversion of Amplex Red to resorufin requires the reaction of horseradish peroxidase with H2O2.

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

View all Réactifs Amplex™ Red et Amplex™ UltraRed FAQs

Citations et références (228)

Citations et références
Abstract
Roles for the human ATP-dependent Lon protease in mitochondrial DNA maintenance.
Authors:Lu B,Yadav S,Shah PG,Liu T,Tian B,Pukszta S,Villaluna N,Kutejová E,Newlon CS,Santos JH,Suzuki CK
Journal:The Journal of biological chemistry
PubMed ID:17420247
Pressure overload-induced mild cardiac hypertrophy reduces left ventricular transmural differences in mitochondrial respiratory chain activity and increases oxidative stress.
Authors:Kindo M, Gerelli S, Bouitbir J, Charles AL, Zoll J, Hoang Minh T, Monassier L, Favret F, Piquard F, Geny B,
Journal:Front Physiol
PubMed ID:22934079
Objective: Increased mechanical stress and contractility characterizes normal left ventricular (LV) subendocardium (Endo) but whether Endo mitochondrial respiratory chain complex activities is reduced as compared to subepicardium (Epi) and whether pressure overload-induced LV hypertrophy (LVH) might modulate transmural gradients through increased reactive oxygen species (ROS) production is unknown. Methods: LVH ... More
Overexpression of nicastrin increases Abeta production.
Authors:Murphy MP, Das P, Nyborg AC, Rochette MJ, Dodson MW, Loosbrock NM, Souder TM, McLendon C, Merit SL, Piper SC, Jansen KR, Golde TE
Journal:FASEB J
PubMed ID:12692078
Gamma-secretase cleavage is the final proteolytic step that releases the amyloid beta-peptide (Abeta) from the amyloid beta-protein precursor (APP). Significant evidence indicates that the presenilins (PS) are catalytic components of a high molecular weight gamma-secretase complex. The glycoprotein nicastrin was recently identified as a functional unit of this complex based ... More
Association of excitatory amino acid transporters, especially EAAT2, with cholesterol-rich lipid raft microdomains: importance for excitatory amino acid transporter localization and function.
Authors:Butchbach ME, Tian G, Guo H, Lin CL
Journal:J Biol Chem
PubMed ID:15187084
In the present study, we investigated the role of membrane cholesterol in the function of glutamate transporters. Depletion of membrane cholesterol by methyl-beta-cyclodextrin resulted in reduced Na(+)-dependent glutamate uptake in primary cortical cultures. Glial glutamate transporter EAAT2-mediated uptake was more sensitive to this effect. Cell surface biotinylation and immunostaining experiments ... More
The characterization of mitochondrial permeability transition in clonal pancreatic beta-cells. Multiple modes and regulation.
Authors:Koshkin V, Bikopoulos G, Chan CB, Wheeler MB
Journal:J Biol Chem
PubMed ID:15231823
Mitochondrial permeability transition (MPT), which contributes substantially to the regulation of normal mitochondrial metabolism, also plays a crucial role in the initiation of cell death. It is known that MPT is regulated in a tissue-specific manner. The importance of MPT in the pancreatic beta-cell is heightened by the fact that ... More
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