Hey everyone,

I'm working on creating a flow QC panel. I have a few sets of 3 intracellular and 3 surface markers that I know have worked well in the past for ICC and flow applications. My intracellular markers are all non-conjugated so I need to use secondary antibodies to put on the fluorochromes. I've titrated the initial unlabelled primary antibodies for the markers but I'm wondering if anybody sees the need to titrate the secondaries. Does anyone have some thoughts on this topic or should I just go with a 1:1000 dilution of them and wash a couple time to remove any unbound secondary?

Thanks,

Mike