LIVE/DEAD™ Fixable Near-IR Dead Cell Stain Kit, for 633 or 635 nm excitation

Citations & References (16)

Invitrogen™

LIVE/DEAD™ Fixable Near-IR Dead Cell Stain Kit, for 633 or 635 nm excitation

Name: LIVE/DEAD™ Fixable Near-IR Dead Cell Stain Kit, for 633 or 635 nm excitation
SKU: L10119

Determines 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.

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Catalog Number	Quantity
L10119	200 Assays
L34975	80 assays
L34976	400 Assays

3 Options

Catalog number L10119

Price (JPY)

83,700

Each

Quantity:

200 Assays

The LIVE/DEAD™ Fixable Near-IR 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 red 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

Stable

The product is sold in tubes with DMSO and in lyophilized flow tubes and plates which require no DMSO. Amine reactive dyes in solution will lose their effectiveness over a short period of time, therefore it is recommended to completely use material once rehydrated.

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 Near-IR 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 Near-IR stain was selected based on its fluorescent properties to provide a bright signal when excited with a red laser. The near-IR fluorescent reactive dye has an excitation maximum of ∼633 nm making it ideal for use with the red or HeNe laser with an emission of ∼775 nm. 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.

Specifications

Cell PermeabilityImpermeant

Cell TypeEukaryotic Cells

DescriptionLIVE/DEAD™ Fixable Near-IR Dead Cell Stain Kit, for 633 or 635 nm excitation

Detection MethodFluorescence

Dye TypeLIVE/DEAD™ FixableNear-IR Dead Cell Stain

FormSolid

FormatTube(s)

Quantity200 Assays

Shipping ConditionRoom Temperature

ColorNear-IR

Emission Wavelength Range775 nm

Excitation Wavelength Range633 nm

For Use With (Application)Viability Assay

For Use With (Equipment)Flow Cytometer

Product LineLIVE/DEAD

Product TypeStain

Unit SizeEach

Contents & Storage

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 60^oC 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 -20^oC 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 Near-IR Dead Cell Stain Kit, for 633 or 635 nm excitation FAQs

Citations & References (16)

Citations & References

Abstract

A retinoic acid--rich tumor microenvironment provides clonal survival cues for tumor-specific CD8(+) T cells.

Authors:Guo Y, Pino-Lagos K, Ahonen CA, Bennett KA, Wang J, Napoli JL, Blomhoff R, Sockanathan S, Chandraratna RA, Dmitrovsky E, Turk MJ, Noelle RJ,

Journal:Cancer Res

PubMed ID:22902413

While vitamin A has been implicated in host resistance to infectious disease, little is known about the role of vitamin A and its active metabolite, retinoic acid (RA) in host defenses against cancer. Here, we show that local RA production within the tumor microenvironment (TME) is increased up to 5-fold ... More

Establishment and characterization of a sustained delayed-type hypersensitivity model with arthritic manifestations in C57BL/6J mice.

Authors:Atkinson SM, Usher PA, Kvist PH, Markholst H, Haase C, Nansen A

Journal:Arthritis Res Ther

PubMed ID:22676339

'Rheumatoid arthritis (RA) is a chronic progressive, inflammatory and destructive autoimmune disease, characterised by synovial joint inflammation and bone erosion. To better understand the pathophysiology and underlying immune mechanisms of RA various models of arthritis have been developed in different inbred strains of mice. Establishment of arthritis models with components ... More

IL-2-dependent adaptive control of NK cell homeostasis.

Authors:Gasteiger G, Hemmers S, Bos PD, Sun JC, Rudensky AY

Journal:J Exp Med

PubMed ID:23650439

'Activation and expansion of T and B lymphocytes and myeloid cells are controlled by Foxp3(+) regulatory T cells (T reg cells), and their deficiency results in a fatal lympho- and myeloproliferative syndrome. A role for T reg cells in the homeostasis of innate lymphocyte lineages remained unknown. Here, we report ... More

Comparative analysis of simian immunodeficiency virus gag-specific effector and memory CD8+ T cells induced by different adenovirus vectors.

Authors:Tan WG, Jin HT, West EE, Penaloza-MacMaster P, Wieland A, Zilliox MJ, McElrath MJ, Barouch DH, Ahmed R,

Journal:J Virol

PubMed ID:23175355

'Adenovirus (Ad) vectors are widely used as experimental vaccines against several infectious diseases, but the magnitude, phenotype, and functionality of CD8(+) T cell responses induced by different adenovirus serotypes have not been compared. To address this question, we have analyzed simian immunodeficiency virus Gag-specific CD8(+) T cell responses in mice ... More

The novel BCR-ABL and FLT3 inhibitor ponatinib is a potent inhibitor of the MDR-associated ATP-binding cassette transporter ABCG2.

Authors:Sen R, Natarajan K, Bhullar J, Shukla S, Fang HB, Cai L, Chen ZS, Ambudkar SV, Baer MR,

Journal:Mol Cancer Ther

PubMed ID:22778153

'Ponatinib is a novel tyrosine kinase inhibitor with potent activity against BCR-ABL with mutations, including T315I, and also against fms-like tyrosine kinase 3. We tested interactions between ponatinib at pharmacologically relevant concentrations of 50 to 200 nmol/L and the MDR-associated ATP-binding cassette (ABC) proteins ABCB1, ABCC1, and ABCG2. Ponatinib enhanced ... More

View all LIVE/DEAD™ Fixable Near-IR Dead Cell Stain Kit, for 633 or 635 nm excitation citations