The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 4 - 8 µg/ml.
Use a concentration of 1.25 µg/ml. Detects a band of approximately 16 kDa (predicted molecular weight: 16 kDa). Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.
FunctionHistone H3-like variant which exclusively replaces conventional H3 in the nucleosome core of centromeric chromatin at the inner plate of the kinetochore. Required for recruitment and assembly of kinetochore proteins, mitotic progression and chromosome segregation. May serve as an epigenetic mark that propagates centromere identity through replication and cell division.
Sequence similaritiesBelongs to the histone H3 family.
DomainThe CATD (CENPA targeting domain) region is responsible for the more compact structure of nucleosomes containing CENPA and is necessary and sufficient to mediate the localization into centromeres.
Post-translational modificationsUbiquitinated (Probable). Interaction with herpes virus HSV-1 ICP0 protein, leads to its degradation by the proteasome pathway. Phosphorylation of Ser-7 by Aurora-A/STK6 and Aurora-B/STK12 during prophase is required for localization of Aurora-A/STK6 and Aurora-B/STK12 at inner centromere and is essential for kinetochore function. Initial phosphorylation during prophase is mediated by Aurora-A/STK6 and is maintained by Aurora-B/STK12.
Cellular localizationNucleus. Chromosome > centromere > kinetochore. Localizes exclusively in the kinetochore domain of centromeres. Occupies a compact domain at the inner kinetochore plate stretching across 2 thirds of the length of the constriction but encompassing only one third of the constriction width and height.