| Stability & Storage | 12 months from date of receipt, -20 °C as supplied. |
Fixable Viability Dye 777 is excited by the Red laser (with an excitation maximum of 750 nm) and has fluorescence emission maximum at 777 nm, Fixable Viability Dye 777 has similar emission to APC-Cy7.
Fixable Viability Dye 777 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 777 Stock Solution for each 1~2 ml of cell suspension/PBS (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 777, 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 Human PBMCs (human peripheral blood mononuclear cells) Dying, the reagent be titrated for optimal performance for different cell types. Stain, fix and permeabilize cells as desired for downstream applications.
For reconstitution, pre-warm the kit to room temperature; add 100 µl of DMSO to one vial of Fixable Viability Dye 777 and mix until fully dissolved. After reconstitution, store aliquots at -20°C. Repeated freezing and thawing are not recommended. Use up within 3 months. Repeated freezing and thawing are not recommended.
Note: We recommend dissolving it in 100 µL of DMSO. The DMSO was overfilled to ensure complete dissolution of the required amount.
Human PBMCs (human peripheral blood mononuclear cells) were stained with Fixable Viability Dye 777 and analyzed before fixation (middle Panels) or after fixation (right Panels). Cells alone, without Fixable Viability Dye 777 staining (left Panels). Flow cytometry and data analysis were performed using BD FACSymphony™ A1 and FlowJo™ software.
