Anti-CENPF antibody (ab5)
Key features and details
- Rabbit polyclonal to CENPF
- Suitable for: ICC/IF, IHC-P, WB
- Reacts with: Human
- Isotype: IgG
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- Ethical standards compliant – production is animal-free
Overview
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Product name
Anti-CENPF antibody
See all CENPF primary antibodies -
Description
Rabbit polyclonal to CENPF -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species ICC/IF HumanIHC-P HumanWB Human
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Immunogen
Fusion protein with C-terminus of CENP-F (Human).
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.
One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.
Learn more here.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles. -
Storage buffer
pH: 7.4
Preservative: 0.05% Sodium azide
Constituents: PBS, 50% Glycerol (glycerin, glycerine) -
Concentration information loading... -
Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Conjugation kits
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Isotype control
Applications
The Abpromise guarantee
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.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
| Application | Species |
|---|---|
| ICC/IF |
Human
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| IHC-P |
Human
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| WB |
Human
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| Application | Abreviews | Notes |
|---|---|---|
| ICC/IF | (5) |
1/400 - 1/750.
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| IHC-P |
Use at an assay dependent concentration.
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| 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.
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| Notes |
|---|
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ICC/IF
1/400 - 1/750. |
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IHC-P
Use at an assay dependent concentration. |
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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. |
Target
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Function
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. -
Involvement in disease
Stromme syndrome -
Sequence similarities
Belongs to the centromere protein F family. -
Developmental stage
Gradually accumulates during the cell cycle, reaching peak levels in G2 and M phase, and is rapidly degraded upon completion of mitosis. -
Post-translational
modificationsHyperphosphorylated during mitosis. -
Cellular localization
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
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Database links
- Entrez Gene: 1063 Human
- Omim: 600236 Human
- SwissProt: P49454 Human
- Unigene: 497741 Human
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Alternative names
- AH antigen antibody
- Cell cycle dependent 350K nuclear protein antibody
- CENF antibody
see all
Images
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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.
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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).
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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)This image is courtesy of Kirk McManus, University of British ColumbiaInterpahse 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.
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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)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.
Datasheets and documents
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SDS download
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Datasheet download
References (70)
ab5 has been referenced in 70 publications.
- 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
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