Special Offers
100% Performance Guaranteed
Key Specifications Table
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| H | WB, PIA | Rb | Affinity Purified | Polyclonal Antibody |
| Description | |
|---|---|
| Catalogue Number | 07-574 |
| Replaces | 04-205 |
| Brand Family | Upstate |
| Trade Name |
|
| Description | Anti-CENP-A Antibody |
| Overview | CENP-A is a variant version of histone H3 found only at centromeres. |
| Alternate Names |
|
| Background Information | Histone H3-like centromeric protein A (UniProt: P49450; also known as Centromere autoantigen A, Centromere protein A, CENP-A) is encoded by the CENPA gene (Gene ID: 1058) in human. CENP-A is a histone H3-like nucleosomal protein that is specifically found in centromeric nucleosomes. It replaces conventional H3 in the nucleosome core of centromeric chromatin and serves as an assembly site for the inner kinetochore. The presence of CENP-A is shown to subtly modify the nucleosome structure, and the way DNA is wrapped around the nucleosome. It gives rise to protruding DNA ends that are less well-ordered, and rigid compared to nucleosomes containing histone H3. Amino acids 39-54 in CENP-A are essential to provide flexibility of DNA ends that protrude from nucleosomes. It also contains a CENP-A targeting domain (CATD; aa 75-116), which is responsible for the more compact structure of nucleosomes containing CENP-A. Its expression varies across the cell cycle with higher levels observed in G2 phase. CENP-A is phosphorylated at serine 68 by CDK1 during early mitosis and this phosphorylation abolishes association with chromatin and centromeres, which prevents interaction with Holliday junction recognition protein (HJURP) and inhibits premature assembly of CENP-A into centromeres. Its phosphorylation at serine 7 by Aurora kinase A or B during prophase is reported to be essential for localization of Aurora kinase A and B at inner centromere and is essential for normal cytokinesis. Its initial phosphorylation during prophase is mediated by Aurora kinase A and is maintained by Aurora kinase B. (Ref.: Orthaus, S., et al. (2008). Chembiochem. 9(1); 77-92; Kunitoku, N., et al. (2003). Dev. Cell. 5(6); 853-864; Zeitlin, SG., et al. (2001). J. Cell. Biol. 155(7); 1147-1157). |
| Product Information | |
|---|---|
| Format | Affinity Purified |
| Control |
|
| Presentation | Purified rabbit polyclonal antibody in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide. |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | Anti-CENP-A, Cat. No. 07-574, is a rabbit polyclonal antibody that detects CENP-A and is tested for use in Western Blotting and Peptide Inhibition Assay and Western Blotting. |
| Key Applications |
|
| Application Notes | Tested applications Peptide Inhibition Assay: Target band detection in acid extract of Colcemid-treated HeLa cells was prevented by pre-blocking of a representative lot with the immunogen peptide. Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user. |
| Biological Information | |
|---|---|
| Immunogen | KLH-conjugated linear peptide corresponding to 14 amino acids from the N-terminal region of human Histone H3-like centromeric protein A (CENP-A). |
| Epitope | N-terminal |
| Host | Rabbit |
| Specificity | This rabbit polyclonal antibody specifically detects Histone H3-like centromeric protein A (CENP-A). It targets an epitope within 14 amino acids from the N-terminal region. |
| Isotype | IgG |
| Species Reactivity |
|
| Species Reactivity Note | Human. |
| Antibody Type | Polyclonal Antibody |
| Entrez Gene Number |
|
| Entrez Gene Summary | Centromeres are the differentiated chromosomal domains that specify the mitotic behavior of chromosomes. CENPA encodes a centromere protein which contains a histone H3 related histone fold domain that is required for targeting to the centromere. CENPA is proposed to be a component of a modified nucleosome or nucleosome-like structure in which it replaces 1 or both copies of conventional histone H3 in the (H3-H4)2 tetrameric core of the nucleosome particle. Alternative splicing results in multiple transcript variants encoding distinct isoforms. |
| Gene Symbol |
|
| Purification Method | Affinity Purified |
| UniProt Number |
|
| UniProt Summary | FUNCTION: SwissProt: P49450 # Histone 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. SIZE: 140 amino acids; 15991 Da SUBUNIT: Forms a nucleosome-like histone octamer containing two molecules each of H2A, H2B, CENPA and H4 assembled in one CENPA-H4 heterotetramer and two H2A-H2B heterodimers. Nucleosomes containing CENPA also contain histone H2A variants such as macroH2A H2AFY and H2A.Z/H2AFZ. The CENPA-H4 heterotetramer is more compact and structurally more rigid than corresponding H3-H4 heterotetramers. Component of the CENPA-NAC complex, at least composed of CENPA, CENPC, CENPH, CENPM, CENPN, CENPT and MLF1IP/CENPU. Interacts directly with herpes virus HSV-1 ICP0 protein. SUBCELLULAR LOCATION: Nucleus. Centromere. Kinetochore. Note=Localizes exclusively in the kinetochore domain of centromeres. DOMAIN: SwissProt: P49450 The CATD (CENPA targeting domain) region is responsible for the more compact structure of nucleosomes containinf CENPA and is necessary and sufficient to mediate the localization into centromeres. PTM: Ubiquitinated (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.DISEASE:SwissProt: P49450 # Antibodies against CENPA are present in sera from patients with autoimmune diseases that developed autoantibodies against centrosomal proteins. SIMILARITY: Belongs to the histone H3 family. |
| Molecular Weight | ~17 kDa observed; 15.99 kDa calculated. Uncharacterized bands may be observed in some lysate(s). |
| Product Usage Statements | |
|---|---|
| Quality Assurance | Evaluated by Western Blotting in acid extract of HeLa cells treated with Colcemid. Western Blotting Analysis: A 1:2,000 dilution of this antibody detected CENP-A in acid extract of HeLa cells treated with Colcemid (50 ng/mL; overnight). |
| Usage Statement |
|
| Storage and Shipping Information | |
|---|---|
| Storage Conditions | Recommended storage: +2°C to +8°C. |
| Packaging Information | |
|---|---|
| Material Size | 200 µL |