A replicated chromosome includes two kinetochores that control chromosome segregation during mitosis. The evolutionarily conserved Centromere Protein-C, CENP-C, is a kinetochore assembly protein. CENP-C is located on the fibers of the kinetochore and constitutes a kinetochore organizing center that tightly associates with DNA. CENP-C is necessary for the formation of a functional centromere, which indicates that CENP-C is important for mitotic progression. In addition, CENP-C is lost from centromeres during herpes simplex virus 1 infection, causing substantial structural changes in the kinetochore, which suggests that the structure of CENP-C is regulated during the cell cycle.
Background References
1. Gopalakrishnan S et al. DNMT3B interacts with constitutive centromere protein CENP-C to modulate DNA methylation and the histone code at centromeric regions. Hum Mol Genet 18:3178-3193 (2009).
2. Gascoigne K E et al. Induced ectopic kinetochore assembly bypasses the requirement for CENP-A nucleosomes. Cell 145:410-422 (2011).
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET7108-24) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
Immunohistochemical analysis of paraffin-embedded mouse spleen tissue with Rabbit anti-CENPC antibody (ET7108-24) at 1/50 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7108-24) at 1/50 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded rat spleen tissue with Rabbit anti-CENPC antibody (ET7108-24) at 1/50 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7108-24) at 1/50 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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