LIVE/DEAD™ Fixable Green Dead Cell Stain Kit, for 488 nm excitation
LIVE/DEAD™ Fixable Green Dead Cell Stain Kit, for 488 nm excitation
Citações e referências (8)
Invitrogen™

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

The LIVE/DEAD™ Fixable Green Dead Cell Stain Kit is used to determine the viability of cells prior to the fixationLeia mais
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Número do catálogoQuantity
L23101200 Assays
L34970400 Assays
L3496980 Assays
Número do catálogo L23101
Preço (BRL)
2.504,94
Each
Adicionar ao carrinho
Quantity:
200 Assays
Preço (BRL)
2.504,94
Each
Adicionar ao carrinho
The LIVE/DEAD™ Fixable Green Dead Cell Stain Kit is used to determine the viability of cells prior to the fixation and permeabilization required for intracellular antibody staining or prior to elimination of biohazardous materials using formaldehyde fixation. This kit has been optimized and validated for use with a blue laser flow cytometer.

• Stable—dyes are freeze dried in separate vials to maintain stability

• Robust—staining pattern is the same before and after fixation

• Bright signal—allows for easy distinction between live/dead cells in single channel

View a selection guide for all fixable viability dyes for flow cytometry.

Stable
Unlike products that are sold in solution, the LIVE/DEAD™ Fixable Green Stain has been conveniently packaged in 40-test vials to help ensure the stability and performance of the dye over time. Amine reactive dyes in solution will lose their effectiveness over a short period of time, therefore it is recommended to completely use the vial once rehydrated. If this is not possible, aliquot the vials in small volumes and store at -80°C, avoiding freeze-thaw cycles.

Robust
Dead cell discriminator stains can lose sensitivity after treatment with fixatives such as formaldehyde or ethanol-based fixation methods required for intracellular phosphorylation studies. The LIVE/DEAD™ Fixable Green Stain is an amine reactive dye that binds covalently to intracellular and extracellular amines, and the staining pattern is preserved following formaldehyde fixation.

Optimal brightness
The LIVE/DEAD™ Fixable Green stain was selected based on its fluorescent properties to provide a bright signal when excited with a blue 488 nm laser. The green-fluorescent reactive dye has an excitation maximum of ∼495 nm making it ideal for use with the blue laser and an emission of ∼520 nm allowing collection in the FITC channel. Since live and dead cells can be discriminated using a single dye and a single channel of a flow cytometer, it is an ideal choice for multi-color experiments.

How it works
In cells with compromised membranes, the dye reacts with free amines both in the cell interior and on the cell surface, yielding intense fluorescent staining. In viable cells, the dye's reactivity is restricted to the cell-surface amines, resulting in less intense fluorescence. The difference in intensity is typically greater than 50-fold between live and dead cells, allowing for easy discrimination.

Colors available
LIVE/DEAD™ Fixable Dead Cell Stains are available in a wide variety of colors to meet your multi-color panel needs.
For Research Use Only. Not for use in diagnostic procedures.
Especificações
Cell PermeabilityImpermeant
Cell TypeEukaryotic Cells
DescriptionLIVE/DEAD™ Fixable Green Dead Cell Stain Kit, for 488 nm excitation
Detection MethodFluorescence
Dye TypeOther Label(s) or Dye(s)
FormSolid
FormatTube(s)
Quantity200 Assays
Shipping ConditionRoom Temperature
SolubilityDMSO (Dimethylsulfoxide)
ColorGreen
Emission520
Excitation Wavelength Range495 nm
For Use With (Application)Viability Assay
For Use With (Equipment)Flow Cytometer
Product LineLIVE/DEAD
Product TypeStain
Unit SizeEach
Conteúdo e armazenamento
Contains 5 vials of LIVE/DEAD™ fixable dead cell stain and 500 μL DMSO.

Store at -20°C.

Frequently asked questions (FAQs)

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 LIVE/DEAD™ Fixable Green Dead Cell Stain Kit, for 488 nm excitation, 200 Assays FAQs

Citações e referências (8)

Citações e referências
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
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