- Datasheet
- Specific References (51)
- Protocols
Overview
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Product nameAnti-CENPF antibody
See all CENPF primary antibodies -
DescriptionRabbit polyclonal to CENPF
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Host speciesRabbit
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SpecificityThis antibody is specific for CENP-F.
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Tested applicationsSuitable for: Flow Cyt, ICC/IF, ICC, IHC-P, WB, IPmore details
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Species reactivityReacts with: Mouse, Human
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Immunogen
Fusion protein with C-terminus of CENP-F (Human).
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Positive control
- WB: Hela whole cell lysate ICC: Hela cells, SK-N-SH cells.
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
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Storage bufferPreservative: 0.05% Sodium azide
Constituents: PBS, 50% Glycerol -
Concentration information loading... -
PurityImmunogen affinity purified
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ClonalityPolyclonal
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IsotypeIgG
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Research areas
Associated products
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Compatible Secondaries
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Conjugation kits
Applications
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.
| Application | Abreviews | Notes |
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| Flow Cyt | 1/200. ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody. |
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| 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. |
Target
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FunctionRequired 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.
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Involvement in diseaseStromme syndrome
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Sequence similaritiesBelongs to the centromere protein F family.
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Developmental stageGradually accumulates during the cell cycle, reaching peak levels in G2 and M phase, and is rapidly degraded upon completion of mitosis.
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Post-translational
modificationsHyperphosphorylated during mitosis. -
Cellular localizationCytoplasm, 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
- Entrez Gene: 108000 Mouse
- 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 ColumbiaHeLa 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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Immunocytochemistry/ Immunofluorescence - Anti-CENPF antibody (ab5)This image is courtesy of an anonymous Abreviewab5 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.
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Western blot - Anti-CENPF antibody (ab5)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?) -
Flow Cytometry - Anti-CENPF antibody (ab5)This image is courtesy of an anonymous Abreviewab5 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
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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.
Protocols
References
This product has been referenced in:
- Pagotto S et al. Hsa-miR-155-5p drives aneuploidy at early stages of cellular transformation. Oncotarget 9:13036-13047 (2018). IF ; Human . Read more (PubMed: 29560129) »
- Li P et al. Genes involved in prostate cancer progression determine MRI visibility. Theranostics 8:1752-1765 (2018). Read more (PubMed: 29556354) »
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