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Invitrogen™ LIVE/DEAD™ Reduced Biohazard Cell Viability Kit #1, green & red fluorescence

Catalog No. L7013
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Catalog No. L7013 Supplier Invitrogen™ Supplier No. L7013
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Reduces risk associated with handling potential biohazards

The LIVE/DEAD Reduced Biohazard Cell Viability Kit No. 1 is a convenient and easy-to-use cell viability kit that is designed to reduce the risk associated with handling potential biohazards such as viral, bacterial or protozoan pathogens. The assay monitors viability as a function of the membrane integrity of the cell. Cells with a compromised membrane that are considered to be dead or dying will stain red, whereas cells with an intact membrane will stain green.

Cell Analysis, Cell Counting and Viability, Cell Counting, Viability, and Cryopreservation, Cell Culture, Cell Viability and Cytotoxicity, Cell Viability, Proliferation and Function, Flow Cytometry, Flow Cytometry Viability and Cytotoxicity Assays, Flow Cytometry of Cellular Processes

Order Info

Shipping Condition: Room temperature

Specifications

Cell Type Eukaryotic Cells
Color Green, Red
Content And Storage Store in freezer at -5°C to -30°C and protect from light.
Description LIVE/DEAD™ Reduced Biohazard Cell Viability Kit #1, green and red fluorescence
Detection Method Fluorescence
For Use With (Application) Viability Assay
For Use With (Equipment) Fluorescence Microscope, Flow Cytometer, Microplate Reader
Product Type Cell Viability Kit
Dye Type Other Label(s) or Dye(s)
Emission 516/617
Excitation Wavelength Range 490 nm
Format Tube(s), 96-well plate, Slide(s)
Product Line LIVE/DEAD
Quantity 1 kit
Shipping Condition Room Temperature
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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.

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.

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.

For Research Use Only. Not for use in diagnostic procedures.