Maybe you could use EdU labelling (it incorporates during S-phase), and count by eye under the microscope the proportion of fluorescent cells.This kit is sold by Invitrogen (Click-it EdU detection kit) It depends what question you are trying to address.
If you look at the principle behind cell cycle analysis, flow cytometry is your best bet. I did look for alternatives a while back but concluded that flow cytometry was, perhaps, the best and the most universally accepted method for cell cycle analysis. All the best.
I agree with that the most universally acepted method is FC. Limitations of the our lab I thought that how can I detect cell cycle. I found an ELISA kit Cell Cycle Human In-Cell ELISA (abcam) , than I will verify my results with cell cycle PCR array (qiagen). Is it a good idea ?
Doing WB for different cell cycle markers is one possibility, but Flow Cytometry is usually needed for publishing your work. If you just want to show that cells can keep entering cell cycle (after a drug treatment), then the clonogenic assay is simple, cheap, and reproducible. Good Luck!
Flow cytommetry using Edu staining is the best in my opinion.
Wester blotting and fluorescent microscopy for cell cycle markers are other options, but lower quality than Edu.
The Qiagen RT-Profiler array doesn't seem to exactly correlate with Edu results in my experience, but I think that could be dependent on RNA quality, extraction method, etc.
There is a kit which requires microscopy and imageJ analysis, which functions similarly to flow cytommetry: http://www.biocolor.co.uk/index.php/assay-kits/cell-clock/
Dear William,
Cell clock assay sounds good. Thank you for your valuable answer.
I would add that using PI/DAPI for cell cycle profiles on flow is cheaper than Edu if you're thinking of flow cytommetry, but want to save money.
- Badges
- Science method