BioLegend
Pacific Blue™ anti-mouse CD40 Antibody
124625
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CD40 is a costimulatory ligand found on antigen presenting cells (B cells, dendritic cells, ect.). I wanted to track CD40 expression on dendritic cells during infection and started by titrating anti-CD40 on lymphocytes from the spleen and lymph nodes of naïve mice to determine staining concentration. Gating on B cells, I saw a small but positive population. While BioLegend suggest a more diluted titration, I find that 1:50 dilution has had the best results.
Flow Cytometry
Spleen or pooled lymph nodes from mice
Titrations (as shown) for 30 minutes at 4C along with antibodies against other markers for gating.
Buffer contained BSA and FcBlock
None
Flow cytometry
I saw comparable results when staining B cells (populations shown are gated B cells) in the spleen and pooled lymph nodes (LN) of naïve mice. There was a small CD40+ population. The best staining was with a 1:50 dilution. I used this dilution for later experiments in infected mice seeing only minor increase in fluorescence compared to FMO controls.
N/A
Works to stain CD40
Doesn't stain brightly.
This antibody does work to track CD40 surface expression. I wouldn’t use it to identify or track maturation of antigen presenting cells.