Anti-CENPA antibody (ab20665)
- Product nameAnti-CENPA antibodySee all CENPA primary antibodies ...
- DescriptionRabbit polyclonal to CENPA
- Tested applicationsIHC-P, WB, ICC/IF more details
- Species reactivityReacts with: Human
Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human CENPA.
- Positive controlThis antibody gave a positive control in HeLa Histone and Calf Thymus Histone preparations. This antibody gave a positive result in IHC in the following FFPE tissue: Human breast adenocarcinoma.
- Storage instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
- Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
- Concentration information loading...
- PurityImmunogen affinity purified
- Clonality Polyclonal
- Research Areas
Our Abpromise guarantee covers the use of ab20665 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||IHC-P: Use a concentration of 1 µg/ml.|
|WB||WB: Use a concentration of 1 µg/ml. Detects a band of approximately 17 kDa (predicted molecular weight: 16 kDa).Can be blocked with CENPA peptide (ab21455).|
|ICC/IF||ICC/IF: Use at an assay dependent concentration.|
- 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.
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.
- CENP A antibodyCENP-A antibodycenpa antibody
- CENPA_HUMAN antibodyCentromere autoantigen A antibodyCentromere protein A 17kDa antibodycentromere protein A antibodycentromeric protein A antibodyHistone H3 like centromeric protein A antibodyHistone H3-like centromeric protein A antibody
Anti-CENPA antibody images
ab20665 at a 1/200 dilution staining paraformaldehyde fixed asynchronous HeLa cells by ICC/IF. The cells were incubated with the antibody overnight and then bound antibody was detected using a Cy3 conjugated goat anti-rabbit antibody. This antibody localizes to centromeres as identified by ACA staining (not shown). Nuclei were visualised with DAPI staining.
This image is courtesy of an Abreview submitted by Kirk McManus.
All lanes : Anti-CENPA antibody (ab20665) at 1 µg/ml
Lane 1 : HeLa Histone lysate at 2.5 µg
Lane 2 : Calf Thymus Histone lysate at 0.5 µg
Lane 3 : Calf Thymus Histone lysate at 0.5 µg with
CENPA peptide (ab21455) at 1 µg/ml
Alexa Fluor Goat polyclonal to Rabbit IgG at 1/10000 dilution
Predicted band size : 16 kDa
Observed band size : 17 kDa (why is the actual band size different from the predicted?)
IHC image of CENPA staining in Human breast adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab20665, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Anti-CENPA antibody (ab20665) at 2 µg/ml +
CENPA protein (His tag) (ab73635) at 1 µg
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed (ab97080) at 1/5000 dilution
developed using the ECL technique
Performed under reducing conditions.
Exposure time : 90 seconds
References for Anti-CENPA antibody (ab20665)
ab20665 has not yet been referenced specifically in any publications.