Acceptable but Poorly Staining Antibody

April 12, 2022

BlueskyReddit
Chemical/Biomedical Engineering
University of Toronto
PhD Candidate

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Company:

BD

Product Name:

Cd80 BUV737

Catalog Number:

612773

Clone Number:

16-10A1

Info:Supplier Page View Product Specs

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We are interested in using cutting edge single cell analysis methods (spectral flow cytometry, scRNAseq, etc) to study the mechanisms responsible for establishing and maintaining immune tolerance.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Mouse thymus, lymph nodes, spleen

Primary Incubation

Block: 1:50 in 100uL at 37C for 30 min

Blocking Agent

Fc Block (aCD16/32)

Secondary Incubation

L/D: Zombie NIR 1:4000 in 100 uL at RT for 20 min

Tertiary Incubation

Surface stain: 1:200 in 100uL at 4C for 30 min

Detection

Spectral flow cytometry (Cytek Aurora)

Results Summary

This antibody weakly stains APCs that are expressing T cell costimulatory molecules (CD80/86). Not possible to separate CD80 high from CD80 intermediate expressing cells, but it is possible to separate positive from negative cells with enough confidence for preliminary use/studies.

DOI or PMID #

N/A

Additional Notes

N/A

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Summary

The Good

Acceptable for assessing if costim molecules are expressed at all.

The Bad

Poor staining of APCs, requires UV channel.

The Bottom Line

Not a great flow cytometry antibody if resolution of CD80 expression with a high dynamic range is required for your study, but ok for simply assessing presence/absence of CD80 expression.

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