1uL 2B4-FITC (BioLegend) Works Great

October 10, 2016

Rheumatology
Vanderbilt University
Postdoctoral Fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

BioLegend

Product Name:

FITC anti-human CD244 (2B4) antibody

Catalog Number:

329505

We built a panel to characterize activating and inhibitory receptors on pTFH cells in untreated chronic HIV+ individuals. We chose 2B4-FITC for our panel because it fit well there in relation to the other conjugates in the panel.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Human PBMC

Primary Incubation

1uL 2B4-FITC in 200uL PBS with 10^6 cells; 15 minutes; room temperature

Blocking Agent

None

Secondary Incubation

None

Tertiary Incubation

None

Detection

BD LSRFortessa

Results Summary

1uL of 2B4-FITC worked great to detect 2B4+ on B cells, monocytes, and CD8+ T cells ex vivo. As expected it also detected rare 2B4+ CD4+ T cells. The staining panel contained 10 other conjugates and at 1uL 2B4-FITC worked great and did not interfere with other channels as measured by FMO controls.

Additional Notes

None

Related Categories

Image Gallery

Summary

The Good

1uL per sample yields great staining; played well in a 10-color fluorescence panel

The Bad

None.

The Bottom Line

We were very happy with how this conjugate worked and are planning to continue using it within the pTFH and other multi-color flow cytometry panels in our lab.

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