your first triple should do; should also include antigens to gate-out other populations that aren't DC but express CD11b/CD8a/MHCII

 Thanks Jeremy, actually I'v been trying not to use only the CD11c as a pan-DC method to gate my cells since some papers are showing that it down regulates after DC activation...Do you usually use CD11c to it?

Hi Rafael, I would suggest you to consider to add an excluding gate like a cocktail antibodies containing anti-CD2 (to exclude NKT and T cells,), anti-B220 (to exclude B cells and plasmacytoid DCs), anti-nk1.1 (if the mouse is C57BL6) or anti-DX5 (if the mouse is BALB/C) to exclude iNKDCs in the same colours. Also, you can add to exclude macrophages out is to use anti-CD64. I recommend you to use to all the antibodies as conjugated with biotin and the you can a bright colour like BV421, PE or PerCP-Cy5.5 or APC conjugated with streptavidin.

Thanks, Daniel. Actually, I did one experiment in which I have used that combination of markers CD11b/CD8a, CD103, and CD317/CD205 to assess MHC II after stimulation with peptides...I did see some effect, and now I think to use a more specific panel to see which cell subset is acting or not. Do you know any good reference to recommend?

Hi Rafael, I have to know a little bit more about your gate strategy. Send me a message with your email that we can chat better.