Wed May 09, 2018 2:37 pm by GregBehbehani
Hi Paul,
We have tested saponin perms and other perm agents with the Pd barcoding method and it should work fine; you'll probably get more barcode signal (similar to if you did barcoding after a methanol perm). The question would be if you plan to perform surface staining before or after using the eBiosciences FoxP3 Staining (fix/perm) Buffer.
If you perform barcoding and surface staining before the fix/perm step then you'll just end up needing to do some sort of fix before barcoding and the subtle barcoding perm (you'll thus effectively have two fix and two perm steps), in this case the barcoding should perform normally. If you do all of your steps after the FoxP3 Staining (fix/perm) Buffer, then your barcoding will likely give you a stronger signal, but any fixation sensitive surface markers will end up looking dimmer due to the fix/perm step. A compromise might be to stain any potentially fixation sensitive markers before fixation and barcoding, then do the fix/perm step, then barcode, then stain the remain surface markers and all intracellular markers. Which to choose depends on why your doing the barcoding and which fix sensitive antigens are important for you. Thus some pilot experiments (as Olivier suggests) would probably be a good idea.
Best of luck,
Greg