Muse® Cell Cycle Assay Kit

Protocol tips

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	- Cells are fixed in cold 70% ethanol for 3 hours at −20°C.

Protocol tips
- Cells are fixed in cold 70% ethanol for 3 hours at −20°C.

Publication protocol

The Muse™ Cell Cycle Assay uses a premixed reagent containing the nuclear DNA intercalating stain propidium iodide (PI) and RNAse A in a proprietary formulation. PI discriminates cells at different stages of the cell cycle, based on the differential DNA content in the presence of RNAse to increase the specificity of DNA staining. The samples were centrifuged at 300× g for 5 min and after removing and discarding the supernatant, an appropriate volume of PBS was added to each tube (1 mL of PBS per 1 × 106 cells). After centrifugation and removing of the supernatant, 1 mL of ice cold 70% ethanol was added to the re-suspending cell pellet in the residual PBS. The tubes were capped and frozen at −20 °C for at least 3 h prior to staining. Ethanol-fixed cells were centrifuged at 300× g for 5 min at room temperature and the pellet was re-suspended in PBS. The cells were centrifuged again at 300× g for 5 min at room temperature, the supernatant was removed and discarded and cell pellet was re-suspended in 200 μL of Muse™ Cell Cycle Reagent and incubated for 30 min at room temperature, in the dark. Cell suspension samples were transferred to a 1.5 mL micro-centrifuge tubes prior to analysis.


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Manufacturer protocol

Download the product protocol from Merck Millipore for Muse® Cell Cycle Assay Kit below.

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