Molecular Probes™ SYTOX™ AADvanced™ Dead Cell Stain Kit

Catalog No.	Quantity
50-113-7611	500 Reactions
50-112-1541	100 Reactions

Catalog No. 50-113-7611 Supplier Molecular Probes™ Supplier No. S10274

Description

SYTOX™ AADvanced™ Dead Cell Stain is a new high-affinity nucleic acid stain for the detection of dead cells and analysis of cell cycle using the common 488 nm blue laser in flow cytometry.

• A new high-affinity nucleic acid stain for the detection of dead cells and analysis of cell cycle using the common 488 nm blue laser in flow cytometry
• The dye is spectrally similar to 7-AAD but with rapid kinetics of uptake and relatively low CVs
• SYTOX™ AADvanced™ Dead Cell Stain penetrates cells more efficiently than 7-AAD providing better separation of live and dead cells
• It can also be used with fixed cells for DNA content analysis when paired with RNAse treatment
• Kits contain a vial(s) of dried dye and anhydrous DMSO providing stable shelf life

Specifications

• Cell Permeability: Impermeant
• Color: Red
• Content And Storage: Contains 5 vials of SYTOX™ AADvanced™ dead cell stain and 500 μL dimethylsulfoxide (DMSO).
  
  Store at ≤-20°C and protect from light.
  
• Description: SYTOX™ AADvanced™ Dead Cell Stain Kit
• Detection Method: Fluorescence
• For Use With (Application): Viability Assay
• For Use With (Equipment): Flow Cytometer
• Product Type: Dead Cell Stain Kit
• Dye Type: SYTOX™ AADvanced™
• Emission: 647
• Excitation Wavelength Range: 546 nm
• Form: Solid
• Format: Tube(s)
• Product Line: Molecular Probes, SYTOX
• Quantity: 500 Reactions
• Shipping Condition: Room Temperature
• Solubility: DMSO (Dimethylsulfoxide)
• Sub Cellular Localization: Nucleus, Nucleic Acids

Frequently Asked Questions (FAQs)

How do SYTO dyes bind to DNA?

The binding mode of SYTO nucleic acid stains is unknown. However, the behavior of these and related nucleic acid dyes suggests the following binding properties:

1.They appear to contact the solvent (suggested by sensitivity to salt, divalent cations, and in particular, SDS) and thus are likely to have contacts in the grooves.
2.All SYTO dyes appear to show some base selectivity and are thus likely to have minor groove contacts.
3.They can be removed from nucleic acid via ethanol precipitation; this characteristic is not shared by ethidium bromide and other intercalators. Likewise, the dyes are not removed from nucleic acid via butanol or chloroform extraction. These extraction methods do remove ethidium bromide from nucleic acid. 4. SYTO binding is not affected by nonionic detergents.
5. SYTO dyes are not quenched by BrdU, so they do not bind nucleic acids in precisely the same way as Hoechst 33342 and DAPI ((4′,6-diamidino-2-phenylindole).

SYBR Green I has shown little mutagenicity on frameshift indicator strains, indicating that it isn't likely to strongly intercalate.

I am using SYTOX AAdvanced as a dead cell stain, but all of my cells are labeling even though I am certain that they are supposed to be alive. These are adherent cells that I have trypsinized. Why am I getting false-dead signals?

SYTOX AAdvanced labels only dead cells because it is a cell impermeant dye. The dye can only enter cells that have a compromised plasma membrane. Trypsinization may cause temporary disruption of the plasma membrane, sufficient to allow staining with a cell impermeant dye. You can reduce the “false-dead” problem by either reducing the amount of trypsin and/or reduce the incubation time for trypsinization or use a gentler dissociation reagent such as TrypLE Express, TrypLESelect reagents, or Versene. After trypsinization, wash well, and if possible, allow a recovery time in normal culture media before staining with any of the SYTOX dyes.

My cell cycle data show a single peak, not a proper cell cycle profile. How can I fix this?

There are several factors that contribute to the quality of the cell cycle profile. Cell number, dye concentration, incubation temperature, incubation time, flow rate (on a traditional flow cytometer utilizing hydrodynamic focusing), total number of cells acquired, elimination of dead cells, and removal of aggregates from data analysis should all be considered when analyzing the cell cycle.

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