I suggest to include to have an intermediate step selecting CD161+/CD127+ on Lin-, CD3- live lymphoid cells. NKP44 appears optional to describe a subset of ILC3+ as mentioned in your protocol.

Is it human blood or tissue? In case of tissue you might need to include a marker to exclude mast cells (which are also CD117+), for example exclude using FcERI or CD203c.

Indeed, you should always use CD127 and CD161 as positive ILC markers. In case you're analyzing blood NKp44 is generally not expressed on ILCs.

I got it. Now I have a question about markers for Innate Lymphoid Cells if used mouse samples.  What should I gate for ILC1, ILC2, ILC3, respectively? 

https://www.biolegend.com/media_assets/literature/images/07-0095-00w.pdf

... a nice overview of ILCs.

Thanks all. I was planning to include CD45, Lin, CD127 and FcERI in the panel (but not CD161 - is this necessary?).

As we are also interested in granulocytes, we were trying to squeeze those into the same panel (there is already a separate panel looking at DCs / T cells). Transcription factors / intracellular cytokines are also on the wish list!

The samples will be human BAL, not many cells so we are limited in terms of the analysis we can perform.

Have you got the percentages for each group of ILC in the blood samples ?

Hi Hugo Farne . It seems that CD161 wasn't always included in the research. Some only use CRTH2 and CD127(IL-7Rα). (I couldn't find FACS antibody for ST2 on the website of Biolegend or BD). I also wonder whether CD161 is necessary for the flow cytometry analysis of ILC2. Do you have a clue?

Hi for all, if I want to study ILC in blood samples which CDs should be used esp. for ILC2 and ILC3 ?