Human CCR7 AF488 Antibody For FACS

Microbiology & Immunology
Columbia University
MD/PhD Student

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Company:

Biolegend

Product Name:

Alexa Fluor 488 anti-human CD197 (CCR7) Antibody

Catalog Number:

353205

Human peripheral blood carries various T cell subsets, whose differentiation status can be characterized through expression of key surface markers, including CCR7 and CD45RA isoforms. Through a combination of surface receptor and transcription factor staining, we are interested in transcription factor expression by memory CD4+ and CD8+ T cells. Our multicolor panels necessitate involve staining for CCR7 to identify naive T cells within the lymphocyte gate. However, staining for CCR7 is notoriously challenging, and several clones require staining at 37C, which introduces high background. This clone and AF488 conjugate was selected based on positive recommendations from other labs regarding stronger separation of signal.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Human PBMCs

Primary Incubation

1:100, 30 min, 4C

Blocking Agent

2% FBS in dPBS; 2mM EDTA

Secondary Incubation

N/A

Tertiary Incubation

N/A

Detection

BD Fortessa

Results Summary

PBMCS were obtained from the peripheral blood of healthy human donors via Ficoll-Hypaque density centrifugation. Cells were then stained to describe steady state subsets. Within the CD3+ CD8+ T cell gate, 4 subsets of cells could be identified based on their expression of CCR7 and CD45RA: Naive, Central Memory, Effector Memory, and TEMRA.

DOI or PMID #

N/A

Additional Notes

N/A

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Summary

The Good

Stronger separation of CCR7+ cells than any other clone tried

The Bad

None

The Bottom Line

Best separation of CCR7+ cells in human samples that we have tried so far.

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