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Epigenetics and Nuclear Signaling Chromosome Structure Centromeres

Anti-CENPF抗体(ab5)

  • Datasheet
  • SDS
Reviews (7)Q&A (2)References (70)

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Immunocytochemistry/ Immunofluorescence - Anti-CENPF antibody (ab5)
  • Immunocytochemistry/ Immunofluorescence - Anti-CENPF antibody (ab5)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CENPF antibody (ab5)
  • Immunocytochemistry/ Immunofluorescence - Anti-CENPF antibody (ab5)
  • Immunocytochemistry/ Immunofluorescence - Anti-CENPF antibody (ab5)
  • Western blot - Anti-CENPF antibody (ab5)
  • Immunocytochemistry/ Immunofluorescence - Anti-CENPF antibody (ab5)

Key features and details

  • Rabbit polyclonal to CENPF
  • Suitable for: ICC/IF, IHC-P, WB
  • Reacts with: Mouse, Human
  • Isotype: IgG

重组抗体的批间可重复性更强

Product image
Anti-CENPF antibody [EPR20406] (ab223847)
  • 研究可靠 —— 各批次间结果一致且可重复
  • 长期批量供应 —— 采用重组技术,可实现快速生产
  • 首次实验即可成功 —— 经过大量验证确认了特异性
  • 符合伦理标准 —— 产品不含动物成分

概述

  • 产品名称

    Anti-CENPF抗体
    参阅全部 CENPF 一抗
  • 描述

    兔多克隆抗体to CENPF
  • 宿主

    Rabbit
  • 经测试应用

    适用于: ICC/IF, IHC-P, WBmore details
  • 种属反应性

    与反应: Mouse, Human
  • 免疫原

    Fusion protein with C-terminus of CENP-F (Human).

  • 常规说明

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

性能

  • 形式

    Liquid
  • 存放说明

    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • 存储溶液

    pH: 7.4
    Preservative: 0.05% Sodium azide
    Constituents: PBS, 50% Glycerol (glycerin, glycerine)
  • Concentration information loading...
  • 纯度

    Immunogen affinity purified
  • 克隆

    多克隆
  • 同种型

    IgG
  • 研究领域

    • Epigenetics and Nuclear Signaling
    • Chromosome Structure
    • Centromeres
    • Epigenetics and Nuclear Signaling
    • Cell cycle
    • Chromosome Structure
    • Centromere

相关产品

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)

应用

Our Abpromise guarantee covers the use of ab5 in the following tested applications.

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

应用 Ab评论 说明
ICC/IF 1/400 - 1/750.
IHC-P Use at an assay dependent concentration.
WB 1/1500. Detects a band of approximately 330 kDa (predicted molecular weight: 330 kDa). Block membranes for 1 hour with 5% nonfat dry milk/TBS-T.

靶标

  • 功能

    Required for kinetochore function and chromosome segregation in mitosis. Required for kinetochore localization of dynein, LIS1, NDE1 and NDEL1. Regulates recycling of the plasma membrane by acting as a link between recycling vesicles and the microtubule network though its association with STX4 and SNAP25. Acts as a potential inhibitor of pocket protein-mediated cellular processes during development by regulating the activity of RB proteins during cell division and proliferation. May play a regulatory or permissive role in the normal embryonic cardiomyocyte cell cycle and in promoting continued mitosis in transformed, abnormally dividing neonatal cardiomyocytes. Interaction with RB directs embryonic stem cells toward a cardiac lineage. Involved in the regulation of DNA synthesis and hence cell cycle progression, via its C-terminus. Has a potential role regulating skeletal myogenesis and in cell differentiation in embryogenesis. Involved in dendritic cell regulation of T-cell immunity against chlamydia.
  • 疾病相关

    Stromme syndrome
  • 序列相似性

    Belongs to the centromere protein F family.
  • 发展阶段

    Gradually accumulates during the cell cycle, reaching peak levels in G2 and M phase, and is rapidly degraded upon completion of mitosis.
  • 翻译后修饰

    Hyperphosphorylated during mitosis.
  • 细胞定位

    Cytoplasm, perinuclear region. Nucleus matrix. Chromosome, centromere, kinetochore. Cytoplasm, cytoskeleton, spindle. Relocalizes to the kinetochore/centromere (coronal surface of the outer plate) and the spindle during mitosis. Observed in nucleus during interphase but not in the nucleolus. At metaphase becomes localized to areas including kinetochore and mitotic apparatus as well as cytoplasm. By telophase, is concentrated within the intracellular bridge at either side of the mid-body.
  • Target information above from: UniProt accession P49454 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 数据库链接

    • Entrez Gene: 1063 Human
    • Entrez Gene: 108000 Mouse
    • Omim: 600236 Human
    • SwissProt: P49454 Human
    • Unigene: 497741 Human
    • 别名

      • AH antigen antibody
      • Cell cycle dependent 350K nuclear protein antibody
      • CENF antibody
      • CENP F antibody
      • CENP F kinetochore protein antibody
      • CENP-F antibody
      • CENPF antibody
      • CENPF kinetochore protein antibody
      • CENPF_HUMAN antibody
      • Centromere protein F 350/400ka antibody
      • Centromere protein F antibody
      • Centromere protein F, 350/400kDa antibody
      • CILD31 antibody
      • Hcp 1 antibody
      • Hcp1 antibody
      • Kinetochore protein CENP F antibody
      • Kinetochore protein CENPF antibody
      • Mitosin antibody
      • PRO1779 antibody
      • STROMS antibody
      see all

    图片

    • Immunocytochemistry/ Immunofluorescence - Anti-CENPF antibody (ab5)
      Immunocytochemistry/ Immunofluorescence - Anti-CENPF antibody (ab5)This image is courtesy of Kirk McManus, University of British Columbia

      HeLa cells were labelled with anti-ACA and anti-CENPF (ab5). ab5 was used at a working dilution of 1/400. This image demonstrates the dramatic increase in fluorescence that occurs late in G2 cells (indicated by arrows). In the final panel DAPI is pseudo-coloured blue, while ACA and CENPF are coloured green and red respectively. 40x magnification.

    • Immunocytochemistry/ Immunofluorescence - Anti-CENPF antibody (ab5)
      Immunocytochemistry/ Immunofluorescence - Anti-CENPF antibody (ab5)

      ICC/IF image of ab5 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab5, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit 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).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CENPF antibody (ab5)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CENPF antibody (ab5)
      Ab5 staining Human normal colon tissue. Staining is localised to nuclear compartment.
      Left panel: with primary antibody at 4 ug/ml. Right panel: isotype control.
      Sections were stained using an automated system DAKO Autostainer Plus , at room temperature: sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
    • Immunocytochemistry/ Immunofluorescence - Anti-CENPF antibody (ab5)
      Immunocytochemistry/ Immunofluorescence - Anti-CENPF antibody (ab5)This image is courtesy of Kirk McManus, University of British Columbia

      Interpahse and Prophase HeLa cells were labelled with anti-ACA and anti-CENPF (ab5). ab5 was used at a working dilution of 1/400. This image emphasizes the redistribution of CENPF from the nuclear matrix during late G2 following entry into the initial stages of mitosis (see the accompanying image). Distinct punctate CENPF patterns proximally located in relation to the centromeres can be seen. In the final panel DAPI is pseudo-coloured blue, while ACA and CENPF are green and red respectively. 100x magnification. 

    • Immunocytochemistry/ Immunofluorescence - Anti-CENPF antibody (ab5)
      Immunocytochemistry/ Immunofluorescence - Anti-CENPF antibody (ab5)This image is courtesy of an anonymous Abreview

      ab5 staining CENPF in Mouse embryonic fibroblast cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.3% Triton X-100 and blocked with 3% BSA for 30 minutes at 23°C. Samples were incubated with primary antibody (1/300 in PBS + 0.1% Triton-X 100 + 1% BSA) for 12 hours at 4°C. An Alexa Fluor® 488-conjugated Goat anti-rabbit IgG polyclonal (1/1000) was used as the secondary antibody.

      See Abreview

    • Western blot - Anti-CENPF antibody (ab5)
      Western blot - Anti-CENPF antibody (ab5)
      All lanes : Anti-CENPF antibody (ab5) at 1/1500 dilution

      Lane 1 : Mitotic HeLa Lysate
      Lane 2 : Asynchronous HeLa Lysate
      Lane 3 : 50gamma Mitotic HeLa Lysate
      Lane 4 : 50gamma Asynchronous HeLa Lysate

      Lysates/proteins at 25 µg per lane.

      Predicted band size: 330 kDa
      Observed band size: 310 kDa
      why is the actual band size different from the predicted?

    • Immunocytochemistry/ Immunofluorescence - Anti-CENPF antibody (ab5)
      Immunocytochemistry/ Immunofluorescence - Anti-CENPF antibody (ab5)This image is courtesy of an Abreview submitted by Dr. Stuart Rulten.

      Paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized A549 (human lung carcinoma cell line) cells stained for CENPF (red) using ab5 at 1/500 dilution in ICC/IF. ). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 555) at 1/500 dilution was used as the secondary antibody.

      See Abreview

    实验方案

    • Recommended WB protocol for ab5

    Click here to view the general protocols

    数据表及文件

    • Datasheet
    • SDS
  • 文献 (70)

    发表研究结果有使用 ab5?请让我们知道,以便我们可以引用本数据表中的参考文章。

    ab5 被引用在 70 文献中.

    • Auckland P  et al. CENP-F stabilizes kinetochore-microtubule attachments and limits dynein stripping of corona cargoes. J Cell Biol 219:N/A (2020). PubMed: 32207772
    • Stiff T  et al. Prophase-Specific Perinuclear Actin Coordinates Centrosome Separation and Positioning to Ensure Accurate Chromosome Segregation. Cell Rep 31:107681 (2020). PubMed: 32460023
    • Shen T  et al. XBP1 negatively regulates CENPF expression via recruiting ATF6a to the promoter during ER stress. Cancer Cell Int 20:459 (2020). PubMed: 32973403
    • Dwivedi D  et al. The dynein adaptor Hook2 plays essential roles in mitotic progression and cytokinesis. J Cell Biol 218:871-894 (2019). PubMed: 30674580
    • Martinez-Macias MI  et al. FUS (fused in sarcoma) is a component of the cellular response to topoisomerase I-induced DNA breakage and transcriptional stress. Life Sci Alliance 2:N/A (2019). PubMed: 30808650
    View all Publications for this product

    客户评价及客户问答

    Show All 评价 Q&A
    提交评价 提交问题

    1-9 of 9 Abreviews or Q&A

    Immunocytochemistry/ Immunofluorescence abreview for Anti-CENPF antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 23°C
    Sample
    Mouse Cell (Mouse embryonic fibroblast cells)
    Specification
    Mouse embryonic fibroblast cells
    Permeabilization
    Yes - 0.3% Triton X-100
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    提交于 Oct 15 2013

    Immunocytochemistry/ Immunofluorescence abreview for Anti-CENPF antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (Human lung adenocarcinoma A549)
    Specification
    Human lung adenocarcinoma A549
    Fixative
    Paraformaldehyde
    Permeabilization
    Yes - 0.2% Triton in PBS
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 20°C
    Read More

    Dr. Stuart Rulten

    Verified customer

    提交于 Jan 02 2013

    Question

    I am looking for a positive control for use in WB and I notice that your image shows a strong band in mitotic cells. Can you tell me how these cells were prepared to ensure that they are all mitotic?

    Read More

    Abcam community

    Verified customer

    Asked on Jul 11 2012

    Answer

    Thank you for contacting us.

    While I was unable to locate any information regarding the specific protocol used to synchronize the mitotic cells in the image on our website (which was provided by a collaborator), the following protocol may be helpful.

    Thymidine-Nocodazole Block:

    HeLa cells were plated at a density of 2x106in 150 mm tissue culture dishes in DMEM with 10%FBS and 100U Pen-Strep
    24 hours after plating, cells were arrested with2mM Thymidine for17-18 hours(S-phase block)
    After Thymidine block: remove Thymidine by washing with 1xPBS
    Add fresh DMEM with 10%FBS and 100U Pen-Strep for 4 hours to release cells.
    Add 100ng/ml Nocodazole to the Media for 12 hours (mitotic block)
    Floating mitotic cells were collected, washed twice in 1X PBS, and replated at a density of 4x106 cells in each 150 cm dish.



    This protocol was used to produce the cells in Figure 1 B of this paper: http://www.molbiolcell.org/content/13/6/1977.long

    I hope this helps, please let me know if you need any additional information or assistance.

    Read More

    Abcam Scientific Support

    回复于 Jul 11 2012

    Immunocytochemistry/ Immunofluorescence abreview for Anti-CENPF antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa cells)
    Specification
    HeLa cells
    Fixative
    Formaldehyde
    Permeabilization
    Yes - 0.1% Triton X 100
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 20°C
    Read More

    Abcam user community

    Verified customer

    提交于 Jan 29 2008

    Flow Cytometry abreview for Anti-CENPF antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Flow Cytometry
    Sample
    Human Cell (T98G glioma cells)
    Specification
    T98G glioma cells
    Fixation
    Ethanol
    Permeabilization
    Yes - Ethanol
    Gating Strategy
    intact cells with >80% and <250% of G1 DNA content
    Read More

    Abcam user community

    Verified customer

    提交于 Jan 29 2008

    Immunocytochemistry/ Immunofluorescence abreview for Anti-CENPF antibody

    Good
    Abreviews
    Abreviews
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa)
    Specification
    HeLa
    Fixative
    Paraformaldehyde
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2
    Read More

    Abcam user community

    Verified customer

    提交于 Aug 15 2006

    Question

    I am interested in using this antibody for immunohistochemistry on mouse tissue, as well as attempting chromatin precipitation assays. The datasheet doesn't specify exactly which region of the human protein was used to make the antibody. Can you inform me of the peptide sequence used to raise the antibody so I can identify the level of conservation in the mouse and assess its likelihood of binding to the mouse protein.

    Read More

    Abcam community

    Verified customer

    Asked on Jan 24 2006

    Answer

    The exact immunogen is proprietary information which was not provided by the supplier unfortunately. I BLASTed the last 200+ residues (our datasheet says that it was made from a bacterial fusion protein from the C-terminus of CENP-F). It shows ~75% homology with fragmented mouse protein. Granted, this is a large span of residues to compare but this is a large protein and this is my best guess for where the epitope resides. I hope that this will already help you and am sorry I couldn't help you more,

    Read More

    Abcam Scientific Support

    回复于 Jan 30 2006

    Immunocytochemistry abreview for Anti-CENPF antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry
    Sample
    Human Cell (HeLa, T98G)
    Specification
    HeLa, T98G
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1%
    Read More

    Dr. Lesley McPhail

    Verified customer

    提交于 Nov 09 2005

    Immunocytochemistry/ Immunofluorescence abreview for Anti-CENPF antibody

    Excellent
    Abreviews
    Abreviews
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa, SK-N-SH and HCT116)
    Specification
    HeLa, SK-N-SH and HCT116
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    提交于 Sep 13 2005

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