Low Staining on Multiple Channels
I have been trying to get a series of CyTOF experiments going, but in every experiment I've done some markers(out of our ~35) have been left abysmally stained. Ie max signal around 10^2. This makes it impossible to discern cell populations and start with actual samples, and not just the validations. Some of this is even occurring on DVS conjugated abs. We are using the Nolan protocols. Some staining is fine, but some is not. I am not sure what could be going wrong? Would keeping the cells at room temp for ~15 min after processing lead to Ab dissociation? What other problems could there be aside from Abs? Clearly, the protocol is being executed okay or else all the markers would be off... I don't have much CyTOF experience so any help would be great!
Thanks so much!