Product: Zombie Violet™ Fixable Viability Kit

Catalog No.: 423113

Sergey Seregin, Postdoc University of Michigan

Good Viability Dye on Pacific Blue™ Channel
Overall:	Product Quality:	Ease of Use:
Good viability dye on Pacific Blue channel for flow cytometry.

Experimental Design

Application:	Flow cytometry
Cells used:	Murine splenocytes, BM cell, lamina propria lympnocytes
Brief Protocol:	1. Prepare single cell suspension of "cells of interest" 2. Use 2-5 million cells/tube (or per well if 96-well plate is used for staining) 3. Centrifuge cells to pellet, wash once with PBS 4. Add Zombie Violet (diluted 1:200 in PBS), incubate 15 min (RT, dark) 5. Add surface antibody mix, incubate 15-20 min (RT, dark) 6. Wash cells 1-2 times with FACS buffer, and either proceed to data acquisition or perform standard fixation/permeabilization protocol follow by intracellular antibody staining.
Results Summary:	We typically get 95% viable cells (which are Zombie Violet negative), which has been routinely confirmed by trypan blue.
Additional Notes:

Data Image Click on the image for full size data.

Publication Status This data has not been published.

Technical Service Notes

Note that the dilution mentioned in step 3 above likely refers to dilution of Zombie AFTER it was initially reconstituted with DMSO. Washing after the Zombie staining step is generally recommended to improve results.

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