Hello everone, I am having a problem compensating cells for one of the investigators here. She is using transplated Wt mouse peritoneal cells. She is staining for B220 (pe-cy5), IgM (pe) and IgD (fitc). Single color controls of the same cells look great and compensate nicely. The problem occurs when the B220 and the IgM are together. We don't see this problem with the same antibodies when used with peritoneal cells from Wt mice or transgenic mice. We also do not see this problem when using bone marrow or splenocytes from the transplanted mice. I have back gated the offending cell population to see if they were possibly dead cells or some other sub population, but they are interspersed pretty evenly throughout the cell population. Has anyone else run into this problem? Any tips? Thanks in advance for any help. Cindy McAllister Children's Hospital Research Institute Columbus, Ohio 43205
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