Overview

  • Product nameAnti-INCENP antibody [58-217]
    See all INCENP primary antibodies
  • Description
    Mouse monoclonal [58-217] to INCENP
  • Tested applicationsSuitable for: ICC/IF, IHC-Pmore details
  • Species reactivity
    Reacts with: Human
    Does not react with: Mouse, Rat
  • Immunogen

    His-tagged human INCENP fragment aa231-298.

Properties

Applications

Our Abpromise guarantee covers the use of ab23956 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
ICC/IF
IHC-P
  • Application notesICC/IF: Use at a concentration of 1 µg/ml.
    IHC-P: 1/200. Perform heat mediated antigen retrieval using citrate buffer pH 6.0 before commencing with IHC staining protocol.


    Not yet tested in other applications.
    Optimal dilutions/concentrations should be determined by the end user.
  • Target

    • FunctionComponent of the chromosomal passenger complex (CPC), a complex that acts as a key regulator of mitosis. The CPC complex has essential functions at the centromere in ensuring correct chromosome alignment and segregation and is required for chromatin-induced microtubule stabilization and spindle assembly. Probably acts through association with AURKB or AURKC. Seems to bind directly to microtubules. Controls the kinetochore localization of BUB1.
    • Sequence similaritiesBelongs to the INCENP family.
    • Cellular localizationChromosome > centromere. Cytoplasm > cytoskeleton > spindle. Nucleus. Chromosome > centromere > kinetochore. Localizes to inner kinetochore. Localizes on chromosome arms and inner centromeres from prophase through metaphase and then transferring to the spindle midzone and midbody from anaphase through cytokinesis. Colocalizes with AURKB at mitotic chromosomes.
    • Information by UniProt
    • Database links
    • Alternative names
      • binds and activates aurora B and C in vivo and in vitro antibody
      • Chromosomal passenger protein antibody
      • INCE_HUMAN antibody
      • INCENP antibody
      • Inner centromere protein antibody
      • Inner centromere protein antigens 135/155kDa antibody
      • Inner centromere protein antigens 135kD 155kD antibody
      • Inner centromere protein INCENP antibody
      see all

    Anti-INCENP antibody [58-217] images

    • ICC/IF image of ab23956 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab23956, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).

    • ab23956 staining INCENP in HeLa cells,  by immunofluorescence.

      optimal antibody dilution: 1mg/ml

      ab23956 staining INCENP in HeLa cells,  by immunofluorescence.

      optimal antibody dilution: 1mg/ml

      optimal fixation protocol: PFA/Triton fixation: 10 min room temperature in 3,7 % PFA diluted in PHEM PEM buffer: 45 mM Hepes pH 6,9, 45 mM Pipes pH 6,9, 5mM MgCl2, 10 mM EGTA) containing 0 ,2% Triton X-100, followed  by 3 washes in PBS.  Also gives a good staining with PFA fixation: 10 min in 3,7 % PFA  at room temperature, 5 min in PBS / 0,1%Triton X-100,  then 3 washes in PBS.

      IF staining was performed following a standard protocol: blocking: 30 min; primary antibody: 1 hr; secondary antibody: 45’. All incubations were at 37 °C in PBS/ 0,1% Tween containing 3% BSA.

      This antibody was also tested using NIH-3T3 cells but it did not provide good results with these cells.

    • ab23956 staining human anal cancer tissue sections by IHC-P.  Sections were formaldehyde fixed and subjected to heat mediated antigen retreival in citrate buffer (pH 6) prior to blocking in a commercially available blocking agent for 20 minutes at 20°C.  The primary anibody was diluted 1/200 and incubated with the sample for 40 minutes at 20°C.  A HRP-conjugated goat anti-mouse antibody was used as the secondary.

      See Abreview

    • ab23956 staining INCENP in siRNA-treated HeLa cells by Immunocytochemistry/ Immunofluorescence.

    References for Anti-INCENP antibody [58-217] (ab23956)

    This product has been referenced in:
    • Kim HS  et al. Functional interplay between Aurora B kinase and Ssu72 phosphatase regulates sister chromatid cohesion. Nat Commun 4:2631 (2013). Read more (PubMed: 24149858) »
    • Vazquez-Martin A  et al. Raptor, a positive regulatory subunit of mTOR complex 1, is a novel phosphoprotein of the rDNA transcription machinery in nucleoli and chromosomal nucleolus organizer regions (NORs). Cell Cycle 10:3140-52 (2011). ICC/IF ; Human . Read more (PubMed: 21900751) »

    See all 2 Publications for this product

    Product Wall

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Immunocytochemistry/ Immunofluorescence
    Sample Human Cell (Colon carcinoma)
    Specification Colon carcinoma
    Fixative Paraformaldehyde
    Permeabilization Yes - 0.5% Triton in PBS
    Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 1%
    Username

    Dr. Eleni Petsalaki

    Verified customer

    Submitted Dec 11 2012

    Abcam has not validated the combination of species/application used in this Abreview.
    Application Western blot
    Sample Human Cell lysate - whole cell (HeLa, U2OS, Jurkat)
    Loading amount 20 µg
    Specification HeLa, U2OS, Jurkat
    Gel Running Conditions Reduced Denaturing (5% SDS-PAGE)
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
    Username

    Mr. Antibody Solutions

    Verified customer

    Submitted Aug 22 2008

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample Human Tissue sections (anal cancer)
    Specification anal cancer
    Fixative Formaldehyde
    Antigen retrieval step Heat mediated - Buffer/Enzyme Used: citrate buffer pH 6.0
    Permeabilization No
    Blocking step Sequential pre-diluted peroxidase (10 min) and protein (10 min) blocks. as blocking agent for 20 minute(s) · Concentration: 100% · Temperature: 20°C
    Username

    Mr. Antibody Solutions

    Verified customer

    Submitted Jul 17 2008

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