Technical Data
C2615-03
CENP-A, phosphorylated (Ser7) (Centromere Potein A)
Description:
Modulation of chromatin structure plays a critical role in the regulation of transcription and replication of the genome in eukaryotes. The nucleosome, made up of four core histone proteins (H2A, H2B, H3, and H4), is the primary building block of chromatin. In addition to the growing number of post-translational histone modifications regulating chromatin structure, cells can also exchange canonical histones with variant histones that can directly or indirectly modulate chromatin structure (1). CENP-A, also known as the chromatin-associated protein CSE4 (capping-enzyme suppressor 4-p), is an essential histone H3 variant that replaces canonical histone H3 in centromeric heterochromatin (2). The greatest divergence between CENP-A and canonical histone H3 occurs in the aminoterminal tail of the protein, which binds linker DNA between nucleosomes and facilitates proper folding of centromeric heterochromatin (3). The amino-terminal tail of CENP-A is also required for recruitment of other centromeric proteins (CENP-C, hSMC1, hZW10), proper kinetochore assembly and chromosome segregation during mitosis (4). Additional sequence divergence in the histone fold domain is responsible for correct targeting of CENP-A to the centromere (5). Many of the functions of CENP-A are regulated by phosphorylation (6,7). Aurora-A-dependent phosphorylation of CENP-A on Ser7 during prophase is required for proper targeting of Aurora-B to the inner centromere in prometaphase, proper kinetochore/microtubule attachment and proper alignment of chromosomes during mitosis (6).

Applications:
Suitable for use in Immunofluorescence, Western Blot and Immunoprecipitation. Other applications not tested.

Recommended Dilution:
Western Blot:1:1000
Immunoprecipitation: 1:25
Immunofluorescence: 1:100
Optimal dilutions to be determined by the researcher.

Storage and Stability:
For long-term storage, aliquot and store at -20°C. Aliquots are stable for at least 12 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
TypeIsotypeCloneGrade
PabIgG
SizeStorageShippingSourceHost
100ul-20°CBlue IceHumanRabbit
Concentration:
Immunogen:
Synthetic phosphopeptide (KLH-coupled) corresponding to residues surrounding Ser7 of human CENP-A protein.
Purity:
Purified by peptide affinity chromatography.
Form
Supplied as a liquid in 10mM sodium HEPES, pH 7.5, 150mM sodium chloride, 100ug/ml BSA, 50% glycerol.
Specificity:
Detects endogenous levels of human CENP-A protein only when phosphorylated on Ser7. This antibody does not cross-react with other histone proteins, including Histone H3.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
(1) Jin, J. et al. (2005) Trends Biochem Sci 30, 680–7.
(2) Ausió, J. (2006) Brief Funct Genomic Proteomic 5, 228–43.
(3) Heit, R. et al. (2006) Biochem Cell Biol 84, 605–18.
(4) Van Hooser, A.A. et al. (2001) J Cell Sci 114, 3529–42.
(5) Black, B.E. et al. (2004) Nature 430, 578–82.
(6) Kunitoku, N. et al. (2003) Dev Cell 5, 853–64.
(7) Zeitlin, S.G. et al. (2001) J Cell Biol 155, 1147–57
Orders n 877-616-CELL (2355)