Review of Fixable Viability Dye eFluor 506

Bloodworks NW
Research
Postdoctoral research fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

eBioscience

Product Name:

Fixable Viability Dye eFluor® 506

Catalog Number:

65-0866-14

The Fixable Viability Dye eFluor 506 is a useful dye to distinguish live vs. dead cells prior fixable and permeablization. This dye can be detected in the AmCyan channel. If you are not using any Brilliant Violet 510 conjugated antibody in your panel, this dye is a great option for the AmCyan channel.

Experimental Design and Results Summary

Application

Flow cytometry

Starting Material

Splenocytes

Protocol Overview

1. Cells were first stained with fluorochrome-conjugated antibodies and then washed with PBS. 2. Prepare a master mix of Fixable Fluor 506 by diluting 1ul of the dye with 1ml of PBS for each sample required (ie. for 20 tubes, take 20l of dye + 20ml of PBS). 3. Resuspend pellet with 1ml of diluted Fixable Fluor 506. 4. Incubate at +4C for 20-30 minutes.5. Wash with PBS and ready for analysis on cytometer.

Tips

Read attached user guide from eBioscience carefully to choose the right staining protocol.

Results Summary

Decent separation between live and dead cells. Very easy to use.

Additional Notes

The user manuals for the Fixable dyes from eBioscience are very helpful. Way more helpful and informative than what's provided by Life Technologies.

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Summary

The Good

Good alternative to AmCyan if you need a fixable viability dye.

The Bad

Separation between live and dead cells can be better.

The Bottom Line

Go for it if you are not using any fluorochrome in the AmCyan channel.

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