Guava Cell Cycle Reagent for Flow Cytometry

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- Cells were seeded in serum-free condition for 24 hand made confluent with 50%. - After 24h cells werein grown in a growth medium with or without DHMEQ

Upstream tips
- Cells were seeded in serum-free condition for 24 hand made confluent with 50%. - After 24h cells werein grown in a growth medium with or without DHMEQ

Publication protocol

For cell cycle analysis, cells (50% confluency) were starved in serum-free condition for 24 h, followed by incubation in growth medium with or without DHMEQ. At the end of the treatment, the cells were trypsinized to obtain a single-cell suspension, centrifuged, and the pellet was resuspended in PBS, followed by fixation in ice-cold 70% ethanol. The fixed cells were centrifuged at ×450 g for 5 min, washed with PBS, resuspended in Guava Cell Cycle Reagent (Millipore), incubated for 30 min at room temperature in the dark, and analyzed by Guava EasyCyte flow cytometer (Millipore) according to the manufacturer's instructions. For cell viability assay, cells were seeded into 6-well plates (1 × 105 cells/well) and cultured without or with DHMEQ. The numbers of live cells, apoptotic cells, and dead cells were calculated using Guava ViaCount assay system (Millipore) and Guava EasyCyte flow cytometer with Cytosoft software (Millipore) according to the manufacturer's instructions.

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