Anti-CENPA 抗体 [3-19] - ChIP Grade (ab13939)

製品の概要

  • 製品名Anti-CENPA antibody [3-19] - ChIP Grade
    CENPA 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [3-19] to CENPA - ChIP Grade
  • アプリケーション適用あり: ICC/IF, IHC-P, WB, ChIP, Flow Cytmore details
  • 種交差性
    交差種: Human
    非交差種: Mouse, Rat
  • 免疫原

    Synthetic peptide:

    PRRRSRKPEAPRRRSPS

    , corresponding to amino acids 3-19 of Human CENP A.

  • ポジティブ・コントロール
    • WB: Friend cell lysate ICC: human metaphase chromosomes
  • 特記事項For maximum product recovery centrifuge the product vial before removing cap.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab13939 in the following tested applications.

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

アプリケーション Abreviews 特記事項
ICC/IF Use a concentration of 2 - 10 µg/ml.
IHC-P Use at an assay dependent concentration.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 18 kDa (predicted molecular weight: 16 kDa).
ChIP Use 4-25 µg for µg of chromatin.
Flow Cyt Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

ターゲット情報

  • 機能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.
  • 配列類似性Belongs to the histone H3 family.
  • ドメイン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.
  • 翻訳後修飾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.
  • 細胞内局在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.
  • Information by UniProt
  • 参照データベース
  • 別名
    • 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 [3-19] - ChIP Grade 画像

  • ab13939 staining CENPA in (A) nontumorous liver tissue, and in (B) well-differentiated, (C) moderately-differentiated, and (D) poorly-differentiated HCC. The CENP-A staining is predominantly nuclear in the samples examined, with concurrent diffuse cytoplasmic staining in poorly-differentiated HCC. Primary antibody used at a dilution of 1:500.

  • ab13939 staining CENPA by Immunofluorescence. This antibody works very well in HeLa cells, with double staining of kinetochores using CREST indicating perfect co-localization.

    Optimal antibody dilution: 2mg/ml.

  • Chromatin was prepared from Hela cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10min. The  ChIP was performed with 25µg of chromatin, 4µg of  ab13939 (blue), and 20µl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci). Primers and probes are located in the first kb of the transcribed region.



  • Predicted band size : 16 kDa

    This image is courtesy of Jerome Govin (Grenoble)

    The antibody works well with histones from: Hela cells (human), Friend cells (mouse) and calf thymus (Roche). 5 µg acid-extracted histones were loaded per lane. Images taken following 5 min exposure. 1:1000 dilution.

    Lane 1: Friend cells (mouse)
    Lane 2: Hela cells (human)
    Lane 3: Calf thymus

    The antibody works well with histones from: Hela cells (human), Friend cells (mouse) and calf thymus (Roche). 5 µg acid-extracted histones were loaded per lane. Images taken following 5 min exposure. 1:1000 dilution.

    Lane 1: Friend cells (mouse)

    Lane 2: Hela cells (human)

    Lane 3: Calf thymus

  • Overlay histogram showing HeLA cells stained with ab13939 (red line). The cells were fixed with 100% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab13939, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was Mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

  • This picture shows human metaphase chromosomes detected with ab13939 as primary antibody and AF488 goat anti-mouse secondary antibody (green). This image was kindly supplied as part of the review submitted by Professor Beth A. Sullivan, Boston University School of Medicine.
  • IHC image of ab13939 staining in normal 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 ab13939, 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.

    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.

Anti-CENPA antibody [3-19] - ChIP Grade (ab13939) 使用論文

This product has been referenced in:

See all 45 Publications for this product

Product Wall

Application Western blot
Sample Human Cell lysate - whole cell (HeLa)
Gel Running Conditions Reduced Denaturing (4-15%)
Loading amount 10 µg
Specification HeLa
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Username

Abcam user community

Verified customer

投稿 Sep 08 2015

Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Permeabilization Yes - PBS 0.2% Triton X-100
Specification HeLa
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 23°C
Fixative 4% formaldehyde in PBS 0.2% Triton X-100
Username

Abcam user community

Verified customer

投稿 Aug 07 2015

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (NT2)
Specification NT2
Fixative Paraformaldehyde
Permeabilization Yes - Triton
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C
Username

Mr. Faisal Alzahrani

Verified customer

投稿 Nov 05 2012

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 Methanol
Permeabilization No
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 37°C
Username

Dr. Eleni Petsalaki

Verified customer

投稿 Oct 08 2012

Abcam has not validated the combination of species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Cow Cell (Oocyte, ovarian granulosa cells)
Specification Oocyte, ovarian granulosa cells
Fixative Paraformaldehyde
Permeabilization Yes - Triton X-100
Blocking step BSA as blocking agent for 20 minute(s) · Concentration: 1% · Temperature: 23°C
Username

Prof. Alberto Maria Luciano

Verified customer

投稿 Jan 26 2010

Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (Lymphoblast chromosomes)
Specification Lymphoblast chromosomes
Fixative Carnoy (Methanol: Acetic Acid 3:1)
Permeabilization Yes - 0,25% Triton X-100 in blocking buffer
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: RT°C
Username

Miss. Marie-Chantal Grégoire

Verified customer

投稿 Oct 17 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Fixative Formaldehyde
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

投稿 Dec 11 2007

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (RPE-1)
Specification RPE-1
Fixative Paraformaldehyde
Permeabilization Yes - 0.5% Triton X100 in PBS
Username

Dr. Kirk McManus

Verified customer

投稿 Aug 14 2007

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (HeLa)
Loading amount 75 µg
Specification HeLa
Blocking step Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 3%
Username

Abcam user community

Verified customer

投稿 Feb 19 2007

Thank you for getting back to me. I am sorry that you did not receive a response to your previous enquiry. I appreciate the lengths that you have gone to in order to get this antibody to work, using fresh chromatin. The reduction in signal at the s...

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