Anti-Ki67 抗体 (ab833)

>As I mentioned previously, I would recommend running a no primary antibody control to ensure that the diffuse nuclear staining cannot be attributed to non-specific binding. If the no primary antibody control is clear, then I would conclude thatmost of yourtransfected cells are still proliferating.

Ki67 is not ideal for telling exactly what stage of proliferation your cells are in, butthe first image in this articlemay provide a helpful reference:

http://www.riken.jp/celldynamics/english/research_3.html

In quiescent cells, Ki67 should be entirely absent. For example, you could compare your results to Fig 5 a in this paper: http://genesdev.cshlp.org/content/18/1/88/F5.expansion.htmlor Fig 1 F vs G in this paper: http://www.sciencedirect.com/science/article/pii/S1097276507004042

If you would like to more accurately determine the cell cycle phase of your transfected cells, you could analyze them using propidium iodide (ab14083), DRAQ5 (ab108410),or DRAQ7 (ab109202)in flow cytometry using this protocol:

https://www.abcam.com/index.html?pageconfig=resource&rid=13432

Ki67 will be localized in the perinucleolar region in the G1 phase and in the nuclear matrix (your hazy nuclear stain) in the later S, G2, and M phases. During G0, Ki67 will not be detected. If you are looking to quantify interphase / division vs quiescence / senescence, you can simply look for number of cells showing any fluorescence vs the number of cells without Ki67 expression. In order to make sure that the nuclear staining you are observing is specific, it is always recommended to run a no primary antibody control.