Good Antibody for the Hard-to-Use APC-Cy7 Channel

June 08, 2016

Pediatrics
UW-Madison
Postdoc

Overall

Quality of Results

Ease-of-Optimization

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Company:

eBioscience

Product Name:

Anti-Human/Mouse CD44 APC-eFluor780

Catalog Number:

47-0441-82

CD44 is a good marker of T cell activation. I have used this antibody to look at cytokine producing T cells (in mice). Often APC-conjugated antibodies are not great and can be difficult to use in a large panel; this CD44 APC-eFluor780 antibody is stable (no loss of tandem fluor) and works well even with Alexa700 and APC in the same panel.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Murine lymphocytes (LN)

Primary Incubation

Surface stain with 1:200 dilution, 0-4C for at least 20 minutes (typically incubate for 30 min)

Blocking Agent

NA

Secondary Incubation

NA, ICS after surface stain has no detrimental effect on CD44 antibody stain.

Tertiary Incubation

NA

Detection

Fluorescence

Results Summary

CD44+ cells were easily distinguished from CD44- cells. For the particular experiment CD8+ T cells were investigated and IFNg+ cells were also CD44+ showing that the CD44 antibody worked well. The IFNg antibody for the particular experiment was conjugated to Alexa700, but with compensation there was no issues with the close emission spectra of Alexa700 and APC-eFluor780.

Additional Notes

None

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Summary

The Good

The antibody-fluor combination works well in a difficult flow channel

The Bad

More expensive than other CD44 antibodies, but not for APC conjugates

The Bottom Line

This antibody is worth it for larger panels. I almost always use this conjugate for marking CD44.

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