Hi all,
I am working on the enteric nervous system of zebrafish and I am looking for a cell-cycle marker to label proliferating cells. I have tried antibodies against Ki67 (NCL-Ki67p and 550609) and PCNA (P8825) and none of them worked. I had MCM5 working but the antibody was a gift and we run out. All of them tested with 30min antigen retrieval at 50oC with Citrine buffer, pH6. Any alternatives, or even other protocols/antibodies that could help me?
Thanks a lot!
Cheers,
Elina
This paper (quite old now) did some nice work combining BrdU and Phospho-Rb (Ser795) antibodies I was developing at the time:
Article The perplexed and confused Mutations Affect Distinct Stages ...
It seems CST now has quite a collection of Zebrafish-reactive antibodies, but I imagine they do not routinely test them for staining Zebrafish tissues:
https://www.cellsignal.com/browse/?Nrpp=30&N=4294960393
Phospho-Histone H3 is pretty standard, but will likely just stain mitotic cells, so will be a good indicator of proliferation on the tissue level rather than the cell level:
Article Analysis of Cell Proliferation, Senescence, and Cell Death i...
Thanks a lot for the suggestion Mark. I will try CST, already contacted them :)
Anti-Phospho-histone H3 (ser10) from Millipore (#06-570) has been working great for me to label mitotic cells, both whole-mount and on sections.
Good luck!
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