概述

  • 产品名称
  • 描述
    兔多克隆抗体to CENPF
  • 特异性
    This antibody is specific for CENP-F.
  • 经测试应用
    适用于: Flow Cyt, ICC/IF, ICC, IHC-P, WB, IPmore details
  • 种属反应性
    与反应: Mouse, Human
  • 免疫原

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

  • 阳性对照
    • WB: Hela whole cell lysate ICC: Hela cells, SK-N-SH cells.

性能

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
Flow Cyt 1/200.

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF 1/400 - 1/750.
ICC Use at an assay dependent concentration.
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.
IP 1/50.

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.
  • Information by UniProt
  • 数据库链接
  • 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

Anti-CENPF antibody 图像

  • 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.

  • 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).

  • 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.
  • 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. 

  • 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

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

    Lane 1 : Mitotic
    Lane 2 : Asyn.
    Lane 3 : 50gamma Mitotic HeLa
    Lane 4 : 50gamma Asyn. HeLa lys


    Predicted band size : 330 kDa
    Observed band size : 310 kDa (why is the actual band size different from the predicted?)
  • ab5 staining (human) T98G glioma cells by flow cytometry.  Cells were trypsinized, spun down & resuspended in PBS before fixing and permeabilised in ethanol.  The primary antibody was diluted 1/400 and incubated with the cells for 2 hours at 20°C.  An Alexa Fluor® 488 conjugaetd goat anti-rabit IgG was used as the secondary antibody. 

    Gating strategy: intact cells with >80% and <250% of G1 DNA content

    See Abreview

  • 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

Anti-CENPF antibody (ab5)参考文献

This product has been referenced in:
  • Pfaltzgraff ER  et al. Loss of CENP-F results in distinct microtubule-related defects without chromosomal abnormalities. Mol Biol Cell 27:1990-9 (2016). Read more (PubMed: 27146114) »
  • Eisch V  et al. Progerin impairs chromosome maintenance by depleting CENP-F from metaphase kinetochores in Hutchinson-Gilford progeria fibroblasts. Oncotarget 7:24700-18 (2016). WB, ICC/IF ; Human . Read more (PubMed: 27015553) »

See all 35 Publications for this product

Product Wall

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
Username

Abcam user community

Verified customer

提交于 Oct 15 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Dr. Stuart Rulten

Verified customer

提交于 Jan 02 2013

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 ...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Abcam user community

Verified customer

提交于 Jan 29 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Abcam user community

Verified customer

提交于 Jan 29 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
Paraformaldehyde
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2
Username

Abcam user community

Verified customer

提交于 Aug 15 2006

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% h...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
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%
Username

Dr. Lesley McPhail

Verified customer

提交于 Nov 09 2005

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa, SK-N-SH and HCT116)
Specification
HeLa, SK-N-SH and HCT116
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 Sep 13 2005

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

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