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Products:Epigenetics and Nuclear Signaling >> Histones >> Variants
Overview
Product name
Anti-CENPA antibody
See all CENPA products (10) ...
Description
Rabbit polyclonal to CENPA
Tested applications
IHC-P, WB, ICC/IFmore details
Cross reactivity
Reacts with
Human
Immunogen
Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human CENPA.
Positive control
This antibody gave a positive control in HeLa Histone and Calf Thymus Histone preparations.
Properties
Form
Liquid
Storage instructions
Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage buffer
Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration
Concentration information loading...
Purity
Immunogen affinity purified
Clonality
Polyclonal
Isotype
IgG
Applications
Show applications key
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.
Application notes
IHC-P: 1/100.
WB: Use at 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). Block membrane in 5% BSA for 1 hour. Incubate antibody in TBST.
A previous batch of this rabbit polyclonal antibody was suitable for ICC/IF. However, recent data seem to indicate that the new batch does not perform well for this application. Abcam would recommend the use of ab13939 instead (please bear in mind that this is a mouse monoclonal, not a rabbit polyclonal).
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
Target
Function
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.
Sequence similarities
Belongs to the histone H3 family.
Domain
The 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
modifications
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.
Cellular localization
Nucleus. 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.
Target information above from: UniProt accessionP49450
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).
Alternative names
- CENP A antibody
- CENP-A antibody
- cenpa antibody
see all
Database links
- Entrez Gene: 1058 Human
- Omim: 117139 Human
- SwissProt: P49450 Human
- Unigene: 1594 Human
Anti-CENPA antibody images:
Immunocytochemistry/ Immunofluorescence - CENPA antibody (ab20665)
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.
Western blot - CENPA antibody (ab20665)
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
Secondary
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?)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - CENPA antibody (ab20665)
IHC image of CENPA staining in human Cervical Cancer FFPE section, performed on a 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, 5µg/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
References for Anti-CENPA antibody (ab20665)
ab20665 has not yet been referenced specifically in any publications.
Publishing research using ab20665? Please let us know so that we can cite the reference in this datasheet
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"