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As mitosis is the most defining stage of cell growth, Active Motif's chemiluminescent and colorimetric Mitotic Assay Kits are an easy way to test the effect of various treatments on cellular division. The kits utilize the same highly specific mitosis marker antibody as the fluorescent Mitotic Index Assay, but offer an alternative colorimetric or chemiluminescent readout, enabling you to choose whichever assay format best suits for your needs.
Cell division is a complex, tightly regulated process that is marked by mitosis. Thus, accurate measurement of cells undergoing mitosis is a valuable method to characterize the effects of test compounds on cellular progression.
Specific marker of mitosis
Two significant mitotic events include microtubule spindle formation and chromosome condensation. Histone H3 is phosphorylated on serine 28 during mitotic chromatin condensation before nuclear division occurs, making this event a reliable marker for cells undergoing mitosis.
The Mitotic Assay method
Cells are grown in the included 96-well plates and then treated with whichever compound is being tested for its effect on cell growth. The cells are then fixed and incubated with the highly specific Histone H3 phospho Ser28 monoclonal antibody, followed by an HRP-conjugated secondary antibody for detection. Development is then performed with either colorimetric or chemiluminescent developing reagents. Paclitaxel, which arrests cells in late G2/M phase, is included to create a high-mitotic reference population. Cells can be counted and normalized using the included Crystal Violet stain.
