Anti-CENPA antibody (ab20665)

Overview

• Product nameAnti-CENPA antibodySee all CENPA primary antibodies ...
• Description
  Rabbit polyclonal to CENPA
• Tested applicationsIHC-P, WB, ICC/IF more details
• Species reactivity
  Reacts with: Human
  
• Immunogen

  Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human CENPA.

  Read Abcam's proprietary immunogen policy

• 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.

Properties

• FormLiquid
• 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
• IsotypeIgG
• Research Areas
  • Epigenetics and Nuclear Signaling
  • Cell cycle
  • Chromosome Structure
  • Centromere

  • Epigenetics and Nuclear Signaling
  • Histones
  • Variants

  • Epigenetics and Nuclear Signaling
  • Chromosome Structure
  • Centromeres

Applications

Target

• 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.

Target information above from: UniProt accession P49450 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
• Database links
  • Entrez Gene: 1058 Human
  • Omim: 117139 Human
  • SwissProt: P49450 Human
  • Unigene: 1594 Human

• Alternative names
  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

  see all

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.

  See Abreview

• 

  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) - Anti-CENPA antibody (ab20665)

  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.

• 

  Western blot
  Anti-CENPA antibody (ab20665) at 2 µg/ml + CENPA protein (His tag) (ab73635) at 1 µg
  
  Secondary
  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