Sharp CD8+ T Cell Resolution in Mouse Lymph Node by Flow Cytometry

June 12, 2026

BlueskyReddit
Bioengineering
SII
Principal Investigator

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Company:

BioLegend

Product Name:

Anti-Mouse CD8a Antibody

Catalog Number:

100712

Clone Number:

53-6.7

We used BioLegend’s Anti-Mouse CD8a (clone 53-6.7) to immunophenotype CD8+ cytotoxic T cells in inguinal and mesenteric lymph nodes harvested from C57BL/6 mice. This antibody was part of a multicolor flow cytometry panel aimed at characterizing T cell subsets during a murine infection model.

Experimental Design and Results Summary

Applications

Flow cytometry

Sample

Single-cell suspensions prepared from inguinal and mesenteric lymph nodes of C57BL/6 mice (8–10 weeks old). Lymph nodes were mechanically dissociated through a 70 µm cell strainer, washed in PBS, and resuspended in FACS buffer (PBS + 2% FBS + 2mM EDTA). Cell viability assessed by Zombie Aqua live/dead staining prior to antibody incubation.

Primary Incubation

1:100 dilution (0.5 µg per 1×10⁶ cells) in FACS buffer , 30 minutes at 4°C in the dark

Blocking Agent

TruStain FcX (anti-mouse CD16/32, BioLegend #101320), 10 minutes at 4°C prior to primary antibody

Secondary Incubation

N/A- directly conjugated PE fluorochrome, no secondary required

Tertiary Incubation

N/A

Detection

BD LSRFortessa flow cytometer

Results Summary

Anti-CD8a (53-6.7) yielded a clearly resolved CD8+ population with high signal-to-noise ratio. In inguinal lymph nodes, CD8+ T cells represented ~25% of total live lymphocytes, consistent with published literature. No non-specific staining was observed in FMO controls. The antibody performed robustly across all animals (n=6) with minimal inter-sample variation.

DOI or PMID #

N/A

Additional Notes

N/A

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Summary

The Good

Brilliant resolution of CD8+ T cells straight out of the box. Directly conjugated PE saves time and reduces protocol steps. Highly consistent lot-to-lot. FMO controls were clean and tight.

The Bad

PE conjugate limits flexibility in complex multicolor panels where the PE channel is needed for other markers. A broader range of fluorochrome options would be appreciated.

The Bottom Line

The go-to anti-CD8a clone for mouse flow cytometry, exceptional sensitivity, zero fuss, publication-ready data every time.

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