BrdU labeling for S-phase is easy and well-established (e.g. Yoshinari, N., Ishida, T., Kudo, A., & Kawakami, A. (2009). Gene expression and functional analysis of zebrafish larval fin fold regeneration. Developmental Biology, 325(1), 71–81. http://doi.org/10.1016/j.ydbio.2008.09.028). The more general and better approach would be to get/make a transgenic expressing zFucci in your tissue(s) of interest (see Fukuhara, S., Zhang, J., Yuge, S., Ando, K., Wakayama, Y., Sakaue-Sawano, A., et al. (2014). Visualizing the cell-cycle progression of endothelial cells in zebrafish. Developmental Biology, 393(1), 10–23. http://doi.org/10.1016/j.ydbio.2014.06.015; Kohrman, A. Q., Kim-Yip, R. P., & Posfai, E. (2021). Imaging developmental cell cycles. Biophysical Journal, 120(19), 4149–4161. http://doi.org/10.1016/j.bpj.2021.04.035)

Hi thank you very much for your reply. I realised my question was not clear enough, apologies. I am looking for an antibody for IHCs. The Fucci line is an excellent idea but we don’t have it at the moment and it uses two channels so not very convenient for what I want to do.

thanks again!

Hello Evangelia Fourli

Use proliferating cell nuclear antigen (PCNA). PCNA is present in proliferating cells during every phase of the cell cycle, peaking from G1 to S and decreasing at G2/M, On the other hand, pH3 is a marker of M-phase cells.

I suggest that you may use both PCNA and pH3 expression since PCNA expression can also be detected long after cell cycle exit and can also indicate DNA repair or cell death. If you use PCNA alone as a proliferation marker, it may overestimate the number of cells progressing through the cell cycle.

You may want to refer to the articles attached below for more information.

https://www.biorxiv.org/content/10.1101/2021.06.16.448637v2.full

Article Proliferating cell nuclear antigen (PCNA) as a proliferative...

Best.