I wanted to stain for B-1 Cells in mouse peritoneal cryo samples. I dont know which antibodies should i use for the classification , B220 and CD5 are one which i thought i could use? Does anyone knows the sure/ correct method for the labelling of B1 cells in IHC..
Thank you
seeking your kind assistance...
Your mentioned markers, especially CD5, are good for the labeling of B1 cells in IHC with cryosections. Another good marker for B1 cells is CD43. From the assay point of view, I would suggest you to include appropriate controls (negative control/no-primary control and isotype controls) for confirming the specificity of the staining. You may look at Novus Biologicals' anti-mouse antibodies for these targets : CD5 (clone 53-7.3, catalog# NB100-77359); CD43 (clone R2/60, NBP1-43413); B220 (clone RA3-6B2, catalog# NB100-77420). Good Luck :-)
CD5 is a good marker for identifying B1 cells, but cannot be used on its own because it will label T-cells as well.
I haven't tried this myself, but I would recommend a combination of CD5 and a B-cell-specific transcription factor, e.g., Pax5. This should work in immunohistochemistry because it combines a membrane antigen (CD5) with a nuclear antigen and discrimination of signals should not be a problem.
Good luck
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