AAT Bioquest

Live or Dead™ Fixable Dead Cell Staining Kit *Blue Fluorescence with 405 nm Excitation*


Excitation (nm)
Emission (nm)
Our Live or Dead™ Fixable Dead Cell Staining Kits are a set of tools for labeling cells for fluorescence microscopic investigations of cellular functions. The effective labeling of cells provides a powerful method for studying cellular events in a spatial and temporal context. This particular kit is designed to uniformly label fixed mammalian cells in blue fluorescence for flow cytometry applications with violet laser excitation. The kit uses a proprietary blue fluorescent dye that is more fluorescent upon bonding to cellular components. The fluorescent dye used in the kit is well excited with the violet laser (405 nm excitation) to fluorescence at 460 nm. The kit provides all the essential components with an optimized cell-labeling protocol. It is an excellent tool for preserving of fluorescent images of particular cells, and can also be used for fluorescence flow cytometry applications.


Flow cytometer

Excitation405 nm laser
Emission450/40 nm filter
Instrument specification(s)Pacific Blue channel

Fluorescence microscope

Excitation410 nm
Emission450 nm
Recommended plateBlack wall/clear bottom


Example protocol


Protocol summary

  1. Prepare samples in HHBS (0.5 mL/assay)
  2. Replace with HHBS
  3. Add Stain It™ Violet 450 to the cell suspension
  4. Stain the cells at room temperature or 37°C for 20 - 60 minutes
  5. Wash the cells
  6. Fix the cells (optional)
  7. Examine the sample with flow cytometer and/or fluorescence microscope using the appropriate Excitation/Emission filter 

Important notes
Thaw all the components at room temperature before starting the experiment.


Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles.

1. Stain It™ Violet 450 stock solution (500X):
Add 200 µL DMSO (Component B) into the vial of Stain It™ Violet 450 (Component A) to make 500X Stain It™ Violet 450 stock solution.

For guidelines on cell sample preparation, please visit


Table 1. Fluorescence spectra properties and suggested excitation laser for flow cytometry analysis

Cat. #DescriptionEx (nm)Em (nm)Excitation Source
22500Blue Fluorescence with 405 nm Excitation410450405 nm
22501Green Fluorescence with 405 nm Excitation408512405 nm
22502Orange Fluorescence with 405 nm Excitation398550405 nm
22599Red Fluorescence Optimized for Flow Cytometry523617488 nm
22600Blue Fluorescence353442335 nm
22601Green Fluorescence498521488 nm
22602Orange Fluorescence547573561 nm or 488 nm
22603Red Fluorescence583603 561 nm
22604Deep Red Fluorescence649660633 nm
22605Near Infrared Fluorescence749775633 nm 
  1. Prepare cells using 1X Hanks and 20 mM Hepes buffer (HHBS) or sodium azide-free and serum/protein-free buffer of your choice.

  2. Wash cells once with HHBS or the azide- and serum/protein-free buffer of your choice.

  3. Resuspend cells at 5 - 10 × 106/mL in HHBS or in the azide- and serum/protein-free buffer of your choice.

  4. Add 1 µL of 500X Stain It™ Violet 450 stock solution to 0.5 mL of cells/assay and mix it well.

  5. Incubate at room temperature or 37°C, 5% CO2 incubator for 20 - 60 minutes, protected from light. Note: The optimal stain concentrations and incubation time should be experimentally determined for different cell lines.

  6. Wash cells twice and resuspend cells with HHBS or the buffer of your choice.

  7. Fix cells as desired (optional).

  8. Analyze cells with flow cytometer and/or fluorescence microscope using the appropriate Excitation/Emission filter (see Table 1).


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Spectral properties

Excitation (nm)406
Emission (nm)445



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