Bright and Robust Anti-Human CTLA-4 Staining

December 23, 2023

BlueskyReddit
Charite Strasse 1
Deutsches Rheuma-Forschungszentrum
Research Scientist

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Company:

BD Biosciences

Product Name:

BD Horizon™ BV786 Mouse Anti-Human CD152

Catalog Number:

563931

Clone Number:

BNI3 (RUO)

Info:Supplier Page 

Browse Similar Products: Anti-CD152 Antibody Products

Our research aims to understand how some CD4 T cells become autoreactive in melanoma patients treated with checkpoint inhibitors. In this experiment, we tested the expression of the inhibitory molecule CTLA-4 in CD4 T cells with an anti-CTLA-4 BV786 antibody.

Experimental Design and Results Summary

Applications

Flow cytometry

Sample

Unstimulated and stimulated (PMA+Ionomycin) healthy donor PBMCs

Primary Incubation

Washed twice in PBS and incubated with L/D fixable aqua dye 20 min at 4°C, then washed twice in PBS

Blocking Agent

None

Secondary Incubation

Surface staining with anti-CD4 PE dazzle594, CD3 BUV395, CD45RO BUV737 etc., 30' at 4°C, in the dark, in BD Horizon™ Brilliant Stain Buffer.

Tertiary Incubation

Cells were fixed and permeabilized with commercial kit (FoxpP3 transcription factor kit, eBioscience) for 30 min at 4°C with fixative, 15 min at 4°C with permeabilization buffer, and stained with anti-FOXP3 PerCP-Cy5.5 and anti-CTLA-4 BV786 at 1/80 in permeabilization buffer.

Detection

Violet laser on Symphony cytometer

Results Summary

The antibody identifies CTLA-4 on stimulated cells only. The staining works even with permeabilization for nuclear staining.

DOI or PMID #

N/A

Additional Notes

N/A

Image Gallery

Summary

The Good

Good staining, even after fixation and permeabilization for nuclear staining.

The Bad

Need to be carefully compensated to avoid signal spreading in other channels.

The Bottom Line

Excellent option for CTLA4 detection in human. Not so expensive.

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