Decent Anti-CD127 Antibody From BioLegend for the Flow Cytometric Detection of Memory T Cells

October 16, 2023

Department of Immunology and Microbiology
Scripps Research
Graduate Student

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Company:

BioLegend

Product Name:

Brilliant Violet 421™ anti-human CD127 (IL-7Rα) Antibody

Catalog Number:

351310

Clone Number:

A019D5

The BV 421 CD127 antibody can be used for identifying memory CD8+ T cells within immune cell populations isolated from mouse lungs, spleen, and lymph nodes. The antibody has given good separation at dilution of 1:100 without too many compensation issues. This antibody is known to have very poor affinity so I am not surprised by the high concentration needed for effective staining.

Experimental Design and Results Summary

Applications

Flow cytometry

Sample

Mouse lung, mediastinal lymphnode, spleens

Primary Incubation

1:100, 30 mins at 4C in the dark

Blocking Agent

Fc Shield

Secondary Incubation

BD cytofix/cytoperm; 100 ul for 20 mins at 4 degrees C

Tertiary Incubation

None

Detection

5 laser Aurora Flow cytometer

Results Summary

I used this product to identify memory CD8+ T cells in mice immunized with antigen. Cells were isolated from lungs, Spleen through digestion or smashing through 70 um filters; RBC lysis was performed using ACK buffer. For lungs, Percoll was used for enrichment of lymphocytes. Cells were then stained with viability dye followed by extracellular antibodies such as TCR beta, CD4, CD8, CD127, KLRG1, CD69, CD103.

DOI or PMID #

N/A

Additional Notes

N/A

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Summary

The Good

BV421 is a good fluorophore for this dim antigen.

The Bad

Separation only visible at 1:100 dilution.

The Bottom Line

Given my panel where other bright fluorophores such as PE and APC were already used for tetramers, BV421 conjugated CD127 proved to work well enough to get the job done.

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