Hello. I stained PC12 cells for MAP2 antibody. Both NGF-treated (differentiated) and untreated (undifferentiated - control) PC12 cell stained positive for MAP-2! I used immunocytochemistry and tried different antibodies, but both my control and treated cells expressed MAP2. I expected to see that this marker expressed in PC12 cells only after differentiation but both control and treated cells expressed (stained positive) the marker. Has anyone had the same experience or any suggestion about this matter?
It sounds logical as MAP2 is positive in pheochromocytoma / paraganglioma as well as in other neoplastic and normal tissues of neural crest origin (albeit not in the all of them). What is the biological reason for this protein not to be expressed in PC12 cells?
@Alexander well, PC12 is neural progenitor cell. I expect that the MAP2 does not express as high as it does in PC12 cell that has been treated with NGF and differentiated to neuronal cells.
I would probably check how both your staining technique and primary Ab for MAP2 differ from those previously described. Another option is to perform a cellblock from the culture and compare your previous results with IHC in FFPE sections from the cellblock; however, I do not think it is routinely done.
Thanks Alexander. I am trying different methods now to find out the issue. I was wondering if some one had done the same experiment before, because it is a very common cell model in the study of neuronal cell.
Hi Dear Sara.
I am working on PC12,s and I have gained the same result as you but in realtime PCR.Both groups (diff & cont) have MAP2 exp with a little more exp in diff cells. Id like to know if you have any experiment about it.
Thank you Sara
Wish the best
Hi Shirin. I did some more MAP-2 staining on both differentiated and undifferentiated cells. It was expressed in both, but stronger in differentiated cells.
Hi Dear Sara
thank you. after data analysing , results were significant and up regulated as you said.
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