Anti-CENPA antibody [EP800Y] - Centromere Marker (ab45694)

Overview

  • Product nameAnti-CENPA antibody [EP800Y] - Centromere Marker
    See all CENPA primary antibodies
  • Description
    Rabbit monoclonal [EP800Y] to CENPA - Centromere Marker
  • Tested applicationsSuitable for: WB, IHC-Pmore details
    Unsuitable for: Flow Cyt or ICC/IF
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human CENPA aa 100 to the C-terminus (C terminal).

  • Positive control
    • HeLa cell lysate, Human colon
  • General notes

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated ‘PUR’ on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    Produced using Abcam's RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5, 675, 063 and/or 7, 429, 487.

Properties

Applications

Our Abpromise guarantee covers the use of ab45694 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/1000 - 1/20000. Detects a band of approximately 18 kDa (predicted molecular weight: 16 kDa).
IHC-P 1/250.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

  • Application notesIs unsuitable for Flow Cyt or ICC/IF.
  • 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
      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 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.
    • Information by UniProt
    • Database links
    • Alternative names
      • CENP A antibody
      • CENP-A antibody
      • cenpa antibody
      • CENPA_HUMAN antibody
      • Centromere autoantigen A antibody
      • Centromere protein A 17kDa antibody
      • Centromere protein A antibody
      • Histone H3 like centromeric protein A antibody
      • Histone H3-like centromeric protein A antibody
      see all

    Anti-CENPA antibody [EP800Y] - Centromere Marker images

    • All lanes : Anti-CENPA antibody [EP800Y] - Centromere Marker (ab45694) at 1/1000 dilution (purified)

      Lane 1 : HeLa (human cervix adenocarcinoma) whole cell lysate
      Lane 2 : Human melanoma
      Lane 3 : A375 (human malignant melanoma) whole cell lysate
      Lane 4 : HCT-116 (human colorectal carcinoma) whole cell lysate
      Lane 5 : Human placenta lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

      Predicted band size : 16 kDa
      Observed band size : 18 kDa (why is the actual band size different from the predicted?)
      Blocking and diluting buffer 5% NFDM/TBST
    • Anti-CENPA antibody [EP800Y] - Centromere Marker (ab45694) at 1/20000 dilution (unpurified) + HeLa cell lysate (10ug/lane)

      Predicted band size : 16 kDa
      Observed band size : 18 kDa (why is the actual band size different from the predicted?)
    • Immunohistochemical analysis of paraffin embedded human colon tissue section labelling CENPA with purified ab45694 at dilution of 1/250. The secondary antibody used was HRP-conjugated Goat Anti-Rabbit IgG H&L (ab97051) at dilution of 1/500. The sample was counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset. 

    • IHC image of ab45694 staining CENPA in Human skin 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 unpurified ab45694, 10µg/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.

    • Immunohistochemical analysis of formalin fixed, paraffin embedded colonic adenocarcinoma tissue section labelling CENPA with unpurified ab45694.

    • Immunohistochemical analysis of formalin fixed, paraffin embedded Lymphoma tissue section labelling CENPA with unpurified ab45694.

    • Immunohistochemical analysis of formalin fixed, paraffin embedded prostatic carcinoma tissue section labelling CENPA with unpurified ab45694.

    • Immunohistochemical analysis of formalin fixed, paraffin embedded lung carcinoma tissue section labelling CENPA with unpurified ab45694.

    References for Anti-CENPA antibody [EP800Y] - Centromere Marker (ab45694)

    ab45694 has not yet been referenced specifically in any publications.

    Product Wall

    Abcam has not validated the combination of species/application used in this Abreview.
    Application Immunocytochemistry/ Immunofluorescence
    Sample Human Cell (HeLa)
    Specification HeLa
    Fixative Paraformaldehyde
    Username

    Dr. Kirk McManus

    Verified customer

    Submitted Jul 09 2009

    Thank you for your enquiry. All of the information we have concerning an antibody may be found on the online datasheet. For this particular antibody, the originator has not released an image of the antibody used in IHC to us. However, we do guaran...

    Read More

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"