Post Wed Dec 19, 2018 10:10 am

Combining barcodig with foxp3 staining

We would like to combine surface with Foxp3 and intracellular and Barcoding.
We know that for Foxp3 we will need to have a stronger perm than for barcoding. Has anyone tried to:
- Stain with intracellular cytoplasmic antibodies together with barcoding (after barcoding perm or intracellular Ab perm?)
- Barcoding at the same time than staining with Foxp3 using the stronger perm buffer.
- Barcoding, pooling (at least 10 samples) and then perm deeper for staining with intracellular+Foxp3. In this case should we fix twice of the first fix done for Barcoding is enough also to perm for foxp3?
Thanks a lot for your help!