Amplex™ Red Phospholipase D Assay Kit
Amplex™ Red Phospholipase D Assay Kit
Citas y referencias (17)
Invitrogen™

Amplex™ Red Phospholipase D Assay Kit

El kit de ensayo de fosfolipasa D Amplex™ Red proporciona un método sensible y sencillo para detectar la actividad deMás información
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Número de catálogoCantidad
A12219500 ensayos
Número de catálogo A12219
Precio (MXN)
-
Cantidad:
500 ensayos
El kit de ensayo de fosfolipasa D Amplex™ Red proporciona un método sensible y sencillo para detectar la actividad de fosfolipasa D (PLD) mediante un lector de microplacas de fluorescencia o un fluorímetro.

Consulte nuestra línea completa de ensayos de microplacas de fluorescencia.

• Detecta niveles de actividad de fosfolipasa D de tan solo 10 U/ml
• Su formato permite realizar varias mediciones en un punto de tiempo
• Diseñado para una mínima interferencia de autofluorescencia

La actividad de PC-PLD se supervisa indirectamente usando 10-acetil-3,7-dihidroxifenoxacina (reactivo Amplex™ Red), una sonda fluorogénica sensible para el peróxido de hidrógeno. La PLD convierte la fosfatidilcolina (lecitina) en colina, que luego se oxida mediante la colina oxidasa a betaína y peróxido de hidrógeno. En presencia de la peroxidasa de rábano, el peróxido de hidrógeno reacciona con el reactivo Amplex™ Red en una proporción estequiométrica 1:1 para crear un producto altamente fluorescente, la resorufina.

Como la resorufina tiene unos valores máximos de absorción y emisión de fluorescencia de aproximadamente 571 nm y 585 nm respectivamente, hay poca interferencia de autofluorescencia en la mayoría de las muestras biológicas.

Utilice ensayos Amplex™ Red para una amplia gama de investigaciones
Hay disponible una amplia variedad de ensayos Amplex™ Red para el estudio de la señalización de células y lípidos, la neurobiología, el funcionamiento de procesos inflamatorios e inmunitarios, y el metabolismo. También ofrecemos el reactivo Amplex™ UltraRed (n.º de cat. A36006), un reactivo de segunda generación que proporciona una mayor sensibilidad y fluorescencia más brillante, y el reactivo de parada Amplex™ Red/UltraRed (n.º cat. A33855). El reactivo de parada Amplex™ Red/UltraRed ofrece comodidad y control, ya que permite terminar la reacción de generación de la señal de fluorescencia en un momento determinado por el usuario. Después de añadir el reactivo de parada, la señal de fluorescencia permanece estable durante al menos tres horas. También hay disponibles diseños de ensayos y envases personalizados.
Para uso exclusivo en investigación. No apto para uso en procedimientos diagnósticos.
Especificaciones
Método de detecciónIntensidad de la fluorescencia
Cantidad500 ensayos
Condiciones de envíoTemperatura ambiente
Propiedades de sustratoSustrato a base de lípidos, sustrato químico
Tipo de sustratoSustrato de fosfolipasa
Enzima dianaFosfolipasa
Para utilizar con (aplicación)Ensayo de fosfolipasa D
Para utilizar con (equipo)Lector de microplacas de fluorescencia
Línea de productosAmplex
Tipo de productoEnsayo de fosfolipasa
Unit SizeEach
Contenido y almacenamiento
Almacenar en el congelador de -5 °C a -30 °C y proteger de la luz.

Preguntas frecuentes

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.

Can Amplex Red Assays be performed using cell lysates?

This is not recommended. The presence of endogenous proteases can complicate the assay by degrading the horseradish peroxidase (HRP). Endogenous peroxidases and antioxidants can modify the H2O2 required for the reaction, competing with HRP (and catalase) for the substrate.

The Amplex Red Assays are best performed with either purified enzymes or extracted H2O2 in a defined buffer system, extracellular solutions or body fluids (media, serum, etc.) that do not exhibit high levels of endogenous protease or oxidase activity and do not contain antioxidants.

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

Citations & References (17)

Citations & References
Abstract
Phospholipase D1 production of phosphatidic acid at the plasma membrane promotes exocytosis of large dense-core granules at a late stage.
Authors:Zeniou-Meyer M, Zabari N, Ashery U, Chasserot-Golaz S, Haeberlé AM, Demais V, Bailly Y, Gottfried I, Nakanishi H, Neiman AM, Du G, Frohman MA, Bader MF, Vitale N
Journal:J Biol Chem
PubMed ID:17540765
'Substantial efforts have recently been made to demonstrate the importance of lipids and lipid-modifying enzymes in various membrane trafficking processes, including calcium-regulated exocytosis of hormones and neurotransmitters. Among bioactive lipids, phosphatidic acid (PA) is an attractive candidate to promote membrane fusion through its ability to change membrane topology. To date, ... More
Cholesterol distribution in the Golgi complex of DITNC1 astrocytes is differentially altered by fresh and aged amyloid beta-peptide-(1-42).
Authors:Igbavboa U, Pidcock JM, Johnson LN, Malo TM, Studniski AE, Yu S, Sun GY, Wood WG
Journal:J Biol Chem
PubMed ID:12584199
'The Golgi complex plays an important role in cholesterol trafficking in cells, and amyloid beta-peptides (Abetas) alter cholesterol trafficking. The hypothesis was tested that fresh and aged Abeta-(1-42) would differentially modify Golgi cholesterol content in DINTC1 astrocytes and that the effects of Abeta-(1-42) would be associated with the region of ... More
The antisignaling agent SC-alpha alpha delta 9, 4-(benzyl-(2-[(2,5-diphenyloxazole-4-carbonyl)amino]ethyl)carbamoyl)- 2-decanoylaminobutyric acid, is a structurally unique phospholipid analogue with phospholipase C inhibitory activity.
Authors:Vogt A, Pestell KE, Day BW, Lazo JS, Wipf P,
Journal:Mol Cancer Ther
PubMed ID:12481409
'Phospholipids and lipid second messengers mediate mitogenic signal transduction and oncogenesis, but there have been few successful examples of small molecules that affect biologically important phospholipid metabolism. Here we investigated the actions of a previously described antitumor agent, 4-(benzyl-(2-[(2,5-diphenyloxazole-4-carbonyl)amino]ethyl)carbamoyl)- 2-decanoylaminobutyric acid (SC-alpha alpha delta 9), which has antisignaling properties, on ... More
Interfacial sensing by alveolar type II cells: a new concept in lung physiology?
Authors:Ravasio A, Hobi N, Bertocchi C, Jesacher A, Dietl P, Haller T,
Journal:Am J Physiol Cell Physiol
PubMed ID:21270294
'Alveolar type II (AT II) cells are in close contact with an air-liquid interface (I(AL)). This contact may be of considerable physiological relevance; however, no data exist to provide a satisfying description of this specific microenvironment. This is mainly due to the experimental difficulty to manipulate and analyze cell-air contacts ... More
Loss of the ceramide transfer protein augments EGF receptor signaling in breast cancer.
Authors:Heering J, Weis N, Holeiter M, Neugart F, Staebler A, Fehm TN, Bischoff A, Schiller J, Duss S, Schmid S, Korte T, Herrmann A, Olayioye MA,
Journal:Cancer Res
PubMed ID:22472120
'Triple-negative breast cancers (TNBC) are especially refractory to treatment due to their negative hormone receptor and ErbB2/HER2 status. Therefore, the identification of cancer-associated deregulated signaling pathways is necessary to develop improved targeted therapies. Here, we show that expression of the ceramide transfer protein CERT is reduced in TNBCs. CERT transfers ... More
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