I have been staining NHP samples for flow cytometry analysis with CXCR5 and it seems that the CXCR5+ population is mostly lost when I stain with a panel that includes incubation with BD Cytofix/Cytoperm. However, the population is beautiful when I stain with a panel that only includes surface staining followed by fixation with 1% PFA.

In both stainings, the CXCR5 antibody is included in the surface antibody panel but I suspect that the incubation with BD Cytofix/Cytoperm afterwards is damaging the CXCR5 staining. I have also tried CXCR5 staining at 37C or intracellularly but nothing helped, hence the suspicion about the fixing/permeabilizing buffer.

Has anyone experienced similar problem and figured out a way fo solving it?

Thank you in advance,

Joana Dias

More Joana Dias's questions See All
Similar questions and discussions