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Protein Detection and Quantitation

Mitotic Assay Kits (Color & Chemi)

determine the fold induction of mitosis in a cell population

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.

 
Name Format Cat No. Price  
Mitotic Assay Kit (Color) 2 x 96 rxns 18021 $395 Buy Now
Mitotic Assay Kit (Chemi) 2 x 96 rxns 18022 $445 Buy Now
Histone H3S28ph antibody (mAb) 100 µl 39098 $400 Buy Now

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. The Mitotic Assay Kits include paclitaxel for use as a positive control as it arrests cells in late G2/M phase, which provides a reference point that can be used to compare the effect of other treatments.

 
 
Figure 1: Fold induction of mitosis in HeLa cells.

HeLa cells were seeded at 20,000 cells per well and assayed using the Colorimetric (A) and Chemiluminescent (B) Mitotic Assay Kits. For a reference mitosis population, cells were treated with 1 µM paclitaxel for 6 hours. Results are the average from eight wells.

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.

Contents & Storage

Two 96-well plates for culturing cells, Histone H3 phospho Ser28 monoclonal antibody (Clone HTA28), HRP-conjugated secondary antibody, Paclitaxel, Quenching Solution, 1X Antibody Blocking Buffer, 1X Antibody Dilution Buffer, 10X PBS, 10% Triton X-100, 1% SDS Solution, Developing & Stop Solutions and Crystal Violet Cell Quantification Solution. Storage conditions vary from room temperature to -20°C, see manual for details. All reagents are guaranteed stable for 6 months when stored properly.