Fixable Live/Dead Stain

July 06, 2015

Medicine
Northwestern University
Graduate Student

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Company:

Invitrogen

Product Name:

LIVE/DEAD® Fixable Aqua Dead Cell Stain Kit, for 405 nm excitation

Catalog Number:

L34957

In this experiment we are looking at populations of immune cells in the bone marrow of mice. We use this stain on many cell types to distinguish live cells and dead cells for flow cytometry.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Bone marrow, peritoneal lavage, spleen, lung homogenate

Primary Incubation

0.25 ul in 500 ul PBS for 20 minutes at room temperature

Blocking Agent

Fc block

Secondary Incubation

NA

Tertiary Incubation

NA

Detection

BD LSRII Flow Cytometer

Results Summary

This stain distinguishes live cells from dead cells, but a control sample (like an FMO) must be used in order to properly draw the gate around live cells. We have also used Sytox, which is much better at clearly distinguishing live cells from dead cells. However, Sytox is not a fixable dye. This Aqua live/dead stain is a good option if you need a fixable stain. It is also nice because the Aqua color is rarely used and therefore it is easy to add into a large panel of antibodies.

Additional Notes

None

Related Categories

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Summary

The Good

Fixable dye, color is rarely used and so it is easy to add into a large panel of antibodies

The Bad

FMO is required to draw a gate around the live cells, and even then drawing the gate can be challenging

The Bottom Line

This antibody is a great option for a fixable live/dead stain

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