I am developing a FACs B cell panel for my experiments and will need about 7-8 colours. We use C57BL/6 mice. I am trying to match my fluorochrome brightness and size to the abundance of markers on the cells. My final panel will also be dependent on commercially available antibodies. Currently deciding on what to get and where to get them.
In a WT mice could someone highlight the relative percentages I would expect to see for each marker? Trying to maximise the panel to get the best results as I will likely be using this repeatedly.
Tentatively, my panel:
CD19/B220 APC
IgM FITC
IgD PE
GL7 PE-Cy7
IgG BV421
CD138 APC-Cy7
CXCR4 BV605
Thanks!
Dear Darellynn Oo
Check out one of my old papers on B cell differentiation, maturation and activation. The methods outline all antibody clones and flourochromes (and as we ran almost all experiments on a Canto instrument so we had to restrict our panels to 8 colours). Our knock out mice were done in B6 ES cells/ B6 background so that should help with reference numbers, too. Lastly, you can get most antibodies from Tonbo or Ebioscience, so you don't have to break the bank to do your experiment.
Article MYSM1-dependent checkpoints in B cell lineage differentiatio...
All the best & good luck with your experiment,
Michael
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