Cell Cycle Chromobody® (CCC)
ChromoTek's powerful Cell Cycle Chromobody® enables you to screen compounds such as cancer drugs for effects on the cell cycle and toxicity in one experiment. You can apply this HCA assay in early drug development and validation as well as in basic research.
The Cell Cycle Chromobody® visualizes proliferating cell nuclear antigen (PCNA), the homotrimeric ringshaped protein that encircles the DNA and acts as a stationary loading platform for multiple, transiently interacting factors participating in various DNA transactions. As an essential cellular component, PCNA is highly conserved from yeast to man.
In eukaryotic cells, PCNA plays a key role in DNA replication, repair, cell cycle regulation, and post-replicative transactions like DNA methylation and chromatin remodelling.
Cell Cycle Chromobody® signal during the cell cycle
In G1, the Chromobody® signal is homogeneously distributed throughout the nucleus and cytoplasm. During S phase it accumulates in the nucleus and visualizes the formation of replication foci. In G2, the foci disappear and the cell divides (mitosis).
For simultaneous read out of
- Cell size and morphology
- Nuclear morphology
- Progression of S phase (detailed detection of early-mid-late S phase)
Features of Cell Cycle Chromobody
- Trace dynamic changes during the cell cycle in real time
- Monitor the distribution of an endogenous cell cycle marker protein - in comparison to fluorescent fusion proteins no overexpression artifact or cytotoxic effects
- Does not affect cellular functions
- Available as stable fluorescent HeLa and U2OS cell lines ready to go for cellular screening
Treatment of the U2OS CCC cell line with three exemplary inhibitors of cell cycle progression
16 hrs after addition of increasing concentrations of substances, cells were fixed and DNA counterstained with DAPI. Three independent experiments were performed in duplicate. Image acquisition was performed with an INCell Analyzer 1000 equipped with an 40x air objective. For image analysis and cell classification INCell Workstation software was used to generate a decision tree based on nuclear size, shape and intensity and the amount and size of appearing nuclear granules/ spots (replication foci).