LIVE/DEAD™ Fixable Green Dead Cell Stain Kit, for 488 nm excitation
LIVE/DEAD™ Fixable Green Dead Cell Stain Kit, for 488 nm excitation
Citas y referencias (8)
Invitrogen™

LIVE/DEAD™ Fixable Green Dead Cell Stain Kit, for 488 nm excitation

El kit de tinción de células muertas de fijación LIVE/DEAD™ Green se utiliza para determinar la viabilidad de las célulasMás información
Have Questions?
Cambiar vistabuttonViewtableView
Número de catálogoCantidad
L23101200 ensayos
L34970400 ensayos
L3496980 ensayos
Número de catálogo L23101
Precio (MXN)
-
Cantidad:
200 ensayos
El kit de tinción de células muertas de fijación LIVE/DEAD™ Green se utiliza para determinar la viabilidad de las células antes de la fijación y la permeabilización necesarias para la tinción de anticuerpos intracelulares o antes de la eliminación de materiales biopeligrosos mediante fijación en formaldehído. Este kit se ha optimizado y validado para su uso con un citómetro de flujo de láser azul.

• Estable: los colorantes se han liofilizado en viales separados para mantener la estabilidad

• Sólido: el patrón de tinción es el mismo antes y después de la fijación

• Señal brillante: permite una fácil distinción entre células vivas/muertas en un solo canal
Consulte una guía de selección para todos los colorantes de viabilidad con fijación para citometría de flujo.

Estable
A diferencia de los productos que se venden en forma de solución, la tinción LIVE/DEAD™ Fixable Green se ha envasado cómodamente en viales de 40 ensayos para ayudar a garantizar la estabilidad y el rendimiento del colorante a largo plazo. Al introducirse en la solución, los colorantes de reactivos de amina perderán su eficacia tras un corto periodo de tiempo; por lo tanto, se recomienda utilizar el total del vial una vez que se haya rehidratado. Si esto no es posible, divida los viales en pequeños volúmenes y almacénelos a -80 °C; evite someterlos a ciclos de congelación y descongelación.

Sólido
Los tintes del discriminador de células muertas pueden ver reducida su sensibilidad tras un tratamiento con fijadores, como los métodos de fijación basados en formaldehído o etanol que resultan necesarios para llevar a cabo estudios de fosforilación intracelular. La tinción LIVE/DEAD™ Fixable Green es un tinte de reactivos de amina que se une de forma covalente a las aminas intra y extracelulares. El patrón de tinción se mantiene después de la fijación con formaldehído.

Brillo óptimo
La tinción LIVE/DEAD™ Fixable Green se ha seleccionado basándose en sus propiedades fluorescentes para proporcionar una señal brillante cuando se excita mediante un láser azul de 488 nm. El tinte reactivo fluorescente verde tiene un máximo de excitación de ∼495 nm, por lo que es ideal para su uso con un láser azul y una emisión de ∼520 nm, lo que permite la recopilación en el canal FITC Puesto que se pueden distinguir células vivas y muertas con un solo tinte y un solo canal de un citómetro de flujo, es la opción ideal para experimentos de varios colores.

Funcionamiento
En células con membranas comprometidas, el colorante reacciona con aminas libres tanto en el interior de la célula como en su superficie, lo que provoca una intensa tinción fluorescente. En células viables, la reactividad del colorante está limitada a las aminas de la superficie celular, lo que se traduce en una fluorescencia menos intensa. La diferencia de intensidad suele ser 50 veces mayor entre células vivas y muertas, lo que facilita la discriminación.

Colores disponibles
Los tintes de células muertas con fijación LIVE/DEAD™ están disponibles en una amplia variedad de colores para satisfacer sus necesidades de paneles de varios colores.
Para uso exclusivo en investigación. No apto para uso en procedimientos diagnósticos.
Especificaciones
Permeabilidad celularImpermeable
Tipo de célulaCélulas eucariotas
DescripciónKit de tinción de células muertas LIVE/DEAD™ Fixable Green, para 488 nm de excitación
Método de detecciónFluorescente
Tipo de coloranteOtras etiquetas o colorantes
FormularioSólido
FormatoTubo(s)
Cantidad200 ensayos
Condiciones de envíoTemperatura ambiente
SolubilidadDMSO (dimetilsulfóxido)
ColorVerde
Emission520 nm
Excitation Wavelength Range495 nm
Para utilizar con (aplicación)Ensayo de viabilidad
Para utilizar con (equipo)Citómetro de flujo
Línea de productosLIVE/DEAD
Tipo de productoTinción
Unit SizeEach
Contenido y almacenamiento
Contiene 5 viales de tinción de células muertas con fijación LIVE/DEAD™ y 500 µl de DMSO. Almacenar a -20 °C.

Preguntas frecuentes

I need to use a dead cell control for my viability assay. Do you have a protocol for killing cells for this?

Heat killing is commonly used. Place your cells in a tube in buffer and heat at 60oC for 20 minutes. You can also kill your cells by fixing them with ice cold 70% ethanol for 15 minutes. The ethanol-killed cells can then be stored at -20oC until needed, at which point you wash out the ethanol and replace with buffer.

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

Regarding the LIVE/DEAD Fixable Dead Cell Stain Kits, which can discriminate between live and dead cells using flow cytometry with one emission wavelength. Can these kits be used with microscopy?

This dye gives a dim surface label for live cells, but is internalized and gives a brighter signal for dead cells. Flow cytometry is a very sensitive technique and can easily distinguish between the two populations. Microscopy is not as sensitive and may not be able to distinguish the cells because of a less sensitive detector.

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

How do I prepare dead cell controls for LIVE/DEAD cell viability assays?

There are two easy options. One is to heat-inactivate the cells by placing at 60 degrees C for 20 minutes. The second is to subject the cells to 70% ethanol. Alcohol-fixed cells can be stored indefinitely in the freezer until use, potentially up to several years.

Centrifuge cells, pellet, and remove supernatant.
Fix cells: Add 10 mL ice cold 70% ETOH to a 15 mL tube containing the cell pellet, adding dropwise at first while vortexing, mix well.
Store in freezer until use.
When ready to use, wash twice and resuspend in buffer of choice.

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

Which cell viability kits are compatible with fixation?

The LIVE/DEAD Fixable kits for flow cytometry analysis are compatible with fixation. These kits use amine-reactive cell-impermeant dyes that stain the cell surface of live cells and also the cytosol of dead cells-live cells are dim and dead cells are bright. Since the dye is covalently bound to the cells, it will be retained after fixation. Unfortunately, this method does not work well for imaging-based assays, as all cells are stained and it is difficult to distinguish bright dead cells from dim live cells with a microscope. Ethidium monoazide (EMA; Cat No. E1374) is a cell impermeant nucleic acid stain that can be applied to live cultures and stains only dead cells. After incubation and washing away unbound dye, the cells can be exposed to light to photoactivate EMA to crosslink to dead cell DNA. After crosslinking to dead cell DNA, the samples may be fixed and permeabilized. Image-IT DEAD Green Viability Stain (Cat. No. I10291) for imaging and high-content screening (HCS) analysis is a live-cell impermeant DNA binding dye that is compatible with fixation and permeabilization with good retention up to 48 hours. We also have a LIVE/DEAD Reduced Biohazard Cell Viability Kit (Cat. No. L7013) for imaging and flow analysis that contains two DNA binding dyes, SYTO 10 and Dead Red, that are sufficiently retained to be analyzed soon after 4% glutaraldehyde fixation.
Note: In general, DNA-binding dyes and calcein AM are not compatible with fixation, as these dyes are not covalently bound to components of the cell and will thus slowly diffuse out of cells after fixation, gradually staining all cells as dead.

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

Why do I need to include a viability stain in my assays?

Many antibodies and stains will label dead cells. This will give you misleading data if you do not exclude the dead cells from your analysis. Of course, if you are labeling fixed cells, they are already dead and you do not need a viability stain. However, if you label your cells prior to fixation, then you need to use one of the LIVE/DEAD Fixable Dead Cell Stains.

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

View all Kit de tinción de células muertas LIVE/DEAD™ Fixable Green, para 488 nm de excitación FAQs

Citations & References (8)

Citations & References
Abstract
Techniques to improve the direct ex vivo detection of low frequency antigen-specific CD8+ T cells with peptide-major histocompatibility complex class I tetramers.
Authors:Chattopadhyay PK, Melenhorst JJ, Ladell K, Gostick E, Scheinberg P, Barrett AJ, Wooldridge L, Roederer M, Sewell AK, Price DA,
Journal:Cytometry A
PubMed ID:18836993
'The ability to quantify and characterize antigen-specific CD8+ T cells irrespective of functional readouts using fluorochrome-conjugated peptide-major histocompatibility complex class I (pMHCI) tetramers in conjunction with flow cytometry has transformed our understanding of cellular immune responses over the past decade. In the case of prevalent CD8+ T cell populations that ... More
Amine reactive dyes: an effective tool to discriminate live and dead cells in polychromatic flow cytometry.
Authors:Perfetto SP, Chattopadhyay PK, Lamoreaux L, Nguyen R, Ambrozak D, Koup RA, Roederer M,
Journal:J Immunol Methods
PubMed ID:16756987
'Membrane-damaged cells caused by either mechanical trauma or through normal biological processes can produce artifacts in immunophenotyping analysis by flow cytometry. Dead cells can nonspecifically bind monoclonal antibody conjugates, potentially leading to erroneous conclusions, particularly when cell frequencies are low. To date, DNA intercalating dyes (Ethidium monoazaide (EMA), Propidium Iodide, ... More
Phenotypic Screening with Human iPS Cell-Derived Cardiomyocytes: HTS-Compatible Assays for Interrogating Cardiac Hypertrophy.
Authors:Carlson C, Koonce C, Aoyama N, Einhorn S, Fiene S, Thompson A, Swanson B, Anson B, Kattman S,
Journal:
PubMed ID:24071917
'A major hurdle for cardiovascular disease researchers has been the lack of robust and physiologically relevant cell-based assays for drug discovery. Derivation of cardiomyocytes from human-induced pluripotent stem (iPS) cells at high purity, quality, and quantity enables the development of relevant models of human cardiac disease with source material that ... More
Anakinra treatment in patients with refractory inflammatory myopathies and possible predictive response biomarkers: a mechanistic study with 12 months follow-up.
Authors:Zong M, Dorph C, Dastmalchi M, Alexanderson H, Pieper J, Amoudruz P, Barbasso Helmers S, Nennesmo I, Malmström V, Lundberg IE,
Journal:Ann Rheum Dis
PubMed ID:23625983
'OBJECTIVE: To perform a mechanistic study on the effect of interleukin (IL)-1 blockade by anakinra in patients with refractory myositis and to explore possible predictive biomarkers. METHODS: Fifteen patients with refractory myositis were treated with anakinra for 12 months. Clinical response was assessed by the six-item core set measures of ... More
Blocking angiotensin-converting enzyme induces potent regulatory T cells and modulates TH1- and TH17-mediated autoimmunity.
Authors:Platten M, Youssef S, Hur EM, Ho PP, Han MH, Lanz TV, Phillips LK, Goldstein MJ, Bhat R, Raine CS, Sobel RA, Steinman L,
Journal:Proc Natl Acad Sci U S A
PubMed ID:19706421
The renin-angiotensin-aldosterone system (RAAS) is a major regulator of blood pressure. The octapeptide angiotensin II (AII) is proteolytically processed from the decapeptide AI by angiotensin-converting enzyme (ACE), and then acts via angiotensin type 1 and type 2 receptors (AT1R and AT2R). Inhibitors of ACE and antagonists of the AT1R are ... More
View all Kit de tinción de células muertas LIVE/DEAD™ Fixable Green, para 488 nm de excitación citations