Strongly stains mitotic cells and can therefore be used in flow cytometric analyses of cell suspensions to detect the mitotic index.
Together with a quantitative DNA staining procedure (e.g. propidium iodide) this antibody clearly distinguishes these M-phase cells from cell at other stages of the cell cycle.
Dynamic information can be obtained by combining BrdU incorporation with antibody staining, which can distinguish and quantitate the four major fractions of the cell cycle.
This antibody can be used for flow cytometric analyses and immunocytochemistry, it is not suitable for immunoblotting.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Flow Cyt: 1/50 - 1/100.
ICC: Use at an assay dependent concentration.
IHC-Fr: Use at an assay dependent concentration. Recommended range is 1:50 - 1:100 for immunocytochemistry with avidin-biotinylated horseradish peroxidase complex (ABC) as detection reagent.
Is unsuitable for WB.
This antibody can be used in flow cytometric analyses of cell suspensions to detect the mitotic index.
Using immunocytochemistry, a combination of this antibody and BrdU (ab8955) can distinguish and quantitate the four major fractions of the cell cycle.
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
The life cycle of a eukaryotic cell consists of various phases, two of which can easily be identified. Firstly, during mitosis (M-phase), in which the cell divides into two identical daughter cells, chromosome condensation and spindle formation are microscopically visible. Secondly, in S-phase the DNA of a cell is replicated, a process that can be detected using biochemical techniques. In between the M and S phase two gap phases occur: the G1-phase, the gap between mitosis and the start of DNA replication, and G2-phase, the gap between completion of DNA replication and the onset of mitosis. From G1-phase a cell can leave the cell cycle and enter G0, a ‘quiescent’ phase. Regulation of the cell cycle predominantly occurs at three major control points, which govern the transition from G0 to G1, from G1 to S and from G2 to M-phase.
Delobel P et al. Cell-cycle markers in a transgenic mouse model of human tauopathy: increased levels of cyclin-dependent kinase inhibitors p21Cip1 and p27Kip1. Am J Pathol168:878-87 (2006).
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