Overview

  • Product name
  • Description
    Rabbit polyclonal to INCENP
  • Tested applications
    Suitable for: WB, ICC/IF, ICC, IHC-Fr, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Xenopus laevis
    Predicted to work with: Chicken
  • Immunogen

    Synthetic peptide:

    SKPRYHKRTSSAVWNSP

    (with N-terminal added cysteine) conjugated to KLH, corresponding to amino acids 884-901 of Human INCENP.

  • Positive control
    • Nuclear/whole extract of human HeLa cells.
  • General notes
    If slight turbidity occurs upon prolonged storage, clarify the solution by centrifugation before use. Working dilutions should be discarded if not used within 12 hours. There is some debate about whether this antibody is suitable for use in IF and we would be most interested to hear from researchers who test this antibody in this application. The antibody has been reported to be suitable for use in IF at a dilution of 1/100 using HeLa cells (see image below). However, one of our customers used this product in HeLa cells and reported an unusual staining pattern - please see the enquiries section of the online datasheet for further details.

Properties

Applications

Our Abpromise guarantee covers the use of ab12183 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 1/2000 - 1/5000. Detects a band of approximately 106 kDa (predicted molecular weight: 106 kDa).
ICC/IF Use at an assay dependent concentration.
ICC 1/100.
IHC-Fr Use at an assay dependent concentration.
IP 1/250.

Target

  • Function
    Component 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 similarities
    Belongs to the INCENP family.
  • Cellular localization
    Chromosome > 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 images

  • HeLa cells were fixed for 10mins in fresh 3.7% paraformaldehyde, permeabilised with PBS + 0.1% Triton X 100, and blocked with 1% goat serum in Abdil. The primary antibody (in Abdil) was incubated OVERNIGHT (1 hour didn't work) at room temperature at a dilution of 1/150. The cells were co-stained with mouse anti-tubulin (secondary is Anti-Mouse Alexa 488) and ACA (Human anti centromere marker, secondary is Anti-Human Cy5) and DAPI. The figure shows projected images of dividing cells (prophase, prometaphase, metaphase, anaphase, telophase). The top panels show ab12183, DNA and microtubule stains. The lower panels are the same cells, showing a subset of optical sections at higher magnification. ACA (geen) stains centromeres and ab12183 (red) stains the inner centromere in prophase, prometaphase, and metaphase. The ab12183 channel has been linerally scaled to reduce the background staining associated with this antibody.
  • Demembranated sperm nuclei and CSF (cytostatic factor-arrested) low-speed egg extracts were prepared in XB and reacted at room temperature as described. Proteins were added to extract prior to sperm addition or else immediately after sperm addition into XB buffer (100 mM KCl, 1 mM MgCl2, 0.1 mM CaCl2, 10 mM K-HEPES pH 7.7, 50 mM sucrose) supplemented with energy mix (3.75 mM creatinine phosphate, 0.5 mM Na2-ATP, 0.5 mM MgCl2, 50 µM EGTA). To label damaged DNA, biotin-16-dUTP was added to metaphase reactions to a final concentration of 40 µM. 30 minutes after sperm addition, reactions were diluted 1/10 into XB supplemented with 1 mM MgCl2, 5 mM EGTA, 0.25% Triton X-100, and 10% formaldehyde and processed for Immunofluorescence. Antibody used ab12183 (red).
  • Immunohistochemistry (Frozen sections) analysis of mouse embryonic skin - keratinocytes tissue sections labeling INCENP with ab12183. Tissue was fixed with paraformaldehyde, permeabilized with 10% Triton-X (0.05% Final) and blocked with Gelatin for 1 hour at 20°C. Samples were incubated with primary antibody (1/200) for 12 hours at 4°C. An undiluted Alexa Fluor 488 donkey anti-rabbit IgG polyclonal was used as the secondary antibody.

    See Abreview

References for Anti-INCENP antibody (ab12183)

This product has been referenced in:
  • Pike T  et al. PKC? switches Aurora B specificity to exit the abscission checkpoint. Nat Commun 7:13853 (2016). ICC/IF . Read more (PubMed: 28004745) »
  • Gholkar AA  et al. Fatostatin Inhibits Cancer Cell Proliferation by Affecting Mitotic Microtubule Spindle Assembly and Cell Division. J Biol Chem 291:17001-8 (2016). Read more (PubMed: 27378817) »

See all 12 Publications for this product

Product Wall

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Embryonic Skin - keratinocytes in vivo)
Permeabilization
Yes - 10% Triton-X (0.05% Final)
Specification
Embryonic Skin - keratinocytes in vivo
Blocking step
(agent) for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C
Fixative
Paraformaldehyde
Username

Dr. Scott Williams

Verified customer

Submitted Sep 30 2016

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 22°C
Sample
Human Cell (293T)
Specification
293T
Permeabilization
Yes - 0.5% Triton X100
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Jan 21 2015

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% · Temperature: 37°C
Username

Dr. Eleni Petsalaki

Verified customer

Submitted Dec 03 2012

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa cells)
Specification
HeLa cells
Fixative
Methanol
Permeabilization
Yes - 0.1% triton X-100
Blocking step
BSA as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 4°C
Username

Abcam user community

Verified customer

Submitted Jun 18 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry
Sample
Human Cell (HeLa)
Specification
HeLa
Fixative
Paraformaldehyde
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% in
Username

Dr. William Moore

Verified customer

Submitted Nov 30 2005

Application
Immunocytochemistry
Sample
Human Cell (HeLa and SK-N-SH)
Specification
HeLa and SK-N-SH
Fixative
Paraformaldehyde
Username

Dr. Kirk Mcmanus

Verified customer

Submitted Oct 12 2005

I have received confirmation from the source of ab12183 that the antibody will detect a 106 kDa band using whole HeLa extract. They cannot explain why you are detecting a 80kDa band and hypothesize that this could be a splice variant or a lysed form of...

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We have just received the following information from the source of ab12183 who tested the antibody in HeLa nuclear extracts: 1. Extract: 10^7 cells/slab of HeLa nuclear. 2.Wet transfer for 90 minutes at 4'C. 3. Blocking solution: 5%-10% non fat dr...

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Thanks for your email. I can send you ab2270 as a replacement. There's no need to return ab12183. Can you please confirm your shipping address, including phone number?

Thank you for your enquiry, and I'm sorry to hear that you are experiencing difficulty with ab12183. I would like to make some suggestions in order to assist you. First, I suggest running a positive control. For this antibody nuclear/whole extract of h...

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1-10 of 12 Abreviews or Q&A

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