Fixable Viability Dye 545

Fixable Viability Dye 545 is excited by the Violet laser (with an excitation maximum of 405 nm) and has fluorescence emission maximum at 545 nm, Fixable Viability Dye 545 has similar emission to Brilliant Violet 480™, Amcyan, V500.

Fixable Viability Dye 545 can intracellular diffusion of the dye and covalent binding to higher overall concentrations of amines, each lot of this product is quality control tested, is useful for discrimination of viable from non-viable mammalian cells in multicolor flow cytometric applications.

Each lot of this product is quality control tested. The labeled cells can be fixed with formaldehyde for downstream decontamination, freezing and/or permeabilization and subsequent intracellular staining while maintaining stable viability stain fluorescence.

1.Wash cells one time in PBS buffer.

2.Add 1 µl of Fixable Viability Dye 545 Stock Solution for each 1~2 ml of cell suspension (1:1000~1:2000) and vortex immediately. It is recommended that the reagent be titrated for optimal performance for each application.

3.Resuspend cells at 1*10^7 cells/ml in Fixable Viability Dye 545, Incubate the mixture for 10-15 minutes at room temperature protected from light.

4.Wash cells twice with PBS buffer or equivalent buffer containing serum or BSA.

5.Resuspend the cells in Stain Buffer.

Note: This protocol applicable to mouse splenocytes Dying, the reagent be titrated for optimal performance for different cell types. Stain, fix and permeabilize cells as desired for downstream applications.