Anti-CENPE antibody [1H12] (ab5093)
Key features and details
- Mouse monoclonal [1H12] to CENPE
- Suitable for: ICC/IF, WB, Flow Cyt
- Reacts with: Human
- Isotype: IgG1
Overview
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Product name
Anti-CENPE antibody [1H12]
See all CENPE primary antibodies -
Description
Mouse monoclonal [1H12] to CENPE -
Host species
Mouse -
Tested applications
Suitable for: ICC/IF, WB, Flow Cytmore details -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant full length protein (Human).
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Positive control
- Any human cell line should be suitable as a positive control. Kinetochore staining only visible in mitosis.
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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 this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. 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, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
Preservative: 0.1% Sodium azide
Constituent: PBS -
Concentration information loading...
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Purification notes
Purified from tissue culture supernatant via ion exchange chromatography (>95% total IgG). -
Clonality
Monoclonal -
Clone number
1H12 -
Myeloma
Sp2/0 -
Isotype
IgG1 -
Light chain type
kappa -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab5093 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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ICC/IF | (4) |
Use at an assay dependent concentration.
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WB |
Use a concentration of 0.5 - 1 µg/ml. Predicted molecular weight: 312 kDa. Only suitable for WB if IP is performed first.
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Flow Cyt |
Use 1µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
Notes |
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ICC/IF
Use at an assay dependent concentration. |
WB
Use a concentration of 0.5 - 1 µg/ml. Predicted molecular weight: 312 kDa. Only suitable for WB if IP is performed first. |
Flow Cyt
Use 1µg for 106 cells. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
Target
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Function
Essential for the maintenance of chromosomal stability through efficient stabilization of microtubule capture at kinetochores. Plays a key role in the movement of chromosomes toward the metaphase plate during mitosis. Is a slow plus end-directed motor whose activity is essential for metaphase chromosome alignment. Couples chromosome position to microtubule depolymerizing activity. The highly processive microtubule-dependent motor activity of CENPE serves to power chromosome congression and provides a flexible, motile tether linking kinetochores to dynamic spindle microtubules. Necessary for the mitotic checkpoint signal at individual kinetochores to prevent aneuploidy due to single chromosome loss. Required for the efficient recruitment of BUBR1, MAD1 and MAD2 to attached and newly unattached kinetochores. Stimulates mammalian BUBR1 kinase activity. Accumulates just before mitosis at the G2 phase of the cell cycle. -
Involvement in disease
Microcephaly 13, primary, autosomal recessive -
Sequence similarities
Belongs to the TRAFAC class myosin-kinesin ATPase superfamily. Kinesin family.
Contains 1 kinesin motor domain. -
Domain
The protein is composed of a N-terminal kinesin-motor domain involved in the chromosome movements, a long coil-coiled region involved in the homodimerization and an inhibitory C-tail involved in autoinhibition of the N-terminal catalytic part. -
Post-translational
modificationsThe C-terminal inhibitory domain is phosphorylated. Phosphorylation relieves autoinhibition of the kinetochore motor.
Sumoylated with SUMO2 and SUMO3. The sumoylation mediates the association to the kinetochore. -
Cellular localization
Chromosome, centromere, kinetochore. Cytoplasm, cytoskeleton, spindle. Associates with kinetochores during congression (as early as prometaphase), relocates to the spindle midzone at anaphase, and is quantitatively discarded at the end of the cell division. - Information by UniProt
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Database links
- Entrez Gene: 1062 Human
- Omim: 117143 Human
- SwissProt: Q02224 Human
- Unigene: 75573 Human
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Alternative names
- CENP E antibody
- CENP-E antibody
- CENPE antibody
see all
Images
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Kinetochores specific staining of HCT116 cells arrested in G2/M phase by nocodazole treatment. Methanol fixed cells were stained using mouse monoclonal [1H12] antibody to CENP-E ab5093 (green) and DAPI (blue).
This image was kindly supplied as part of the review submitted by Salvador Rodrigez-Nieto.
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ab5093 at 1/500 staining human fibrosarcoma (HT1080) cells by ICC/IF. The cells were treated with 0.1-0.2ug/mL colcemid for 45-60 minutes, then swollen in hypotonic buffer for 8 minutes and centrifuged onto glass slides. Cells were blocked in 1X PBS + 1% BSA + 0.5% Triton X-100 (blocking buffer) for 30 minutes at room temperature. The antibodies were diluted 1/300-1/500 in blocking buffer and incubated overnight at 4 degrees C. ab5093 was detected with Alexa Fluor 488-donkey anti-mouse for 1-2 hours at room temperature.
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HeLa cells were stained with ab5093, anti-CENPE (in green) in panel one, and with ab5093 and SH-CREST (red), which stains the centromeres, in panel 2. Fix cells for 30 minutes on ice in 4% formaldehyde in PEM. Quench autofluorescence 2 x 5 minutes with 1 mg/ml Na borohydride or 100 mM ammonium chloride in PEM. Permeablize 30 minutes with 0.5% TX-100 in PEM. Block 30 minutes in 5% milk in TBST. Primary antibody overnight at 4oC diluted 1/250 in 5% milk in TBST. Secondary antibody was incubated for 1 hour at RT diluted in 5% milk in TBST. Post-fix 20 minutes on ice in 4% formaldehyde in PEM. Quench autofluorescence 2 x 5 minutes with ammonium chloride in PEM. Counterstain with DAPI in TBST. Mount with ProLong Gold antifade reagent from Invitrogen. Notes: Ample washing between each step. TBST = Tris buffered saline + 0.1% Tween. PEM = 80 mM K-PIPES, pH 6.8, 5 mM EGTA, 2 mM MgCl2.
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All lanes : Anti-CENPE antibody [1H12] (ab5093) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lane 3 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 312 kDa
Observed band size: 270 kDa why is the actual band size different from the predicted?
Exposure time: 20 minutes -
Overlay histogram showing HeLa cells stained with ab5093 (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 (ab5093, 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.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (35)
ab5093 has been referenced in 35 publications.
- Raisch T et al. Structure of the RZZ complex and molecular basis of Spindly-driven corona assembly at human kinetochores. EMBO J 41:e110411 (2022). PubMed: 35373361
- Legal T et al. The C-terminal helix of BubR1 is essential for CENP-E-dependent chromosome alignment. J Cell Sci 133:N/A (2020). PubMed: 32665320
- Waseem NH et al. Mutations in SPATA13/ASEF2 cause primary angle closure glaucoma. PLoS Genet 16:e1008721 (2020). PubMed: 32339198
- Nellikka RK et al. a-Fodrin is required for the organization of functional microtubules during mitosis. Cell Cycle 18:2713-2726 (2019). PubMed: 31455186
- Tan Z et al. Environmental stresses induce karyotypic instability in colorectal cancer cells. Mol Biol Cell 30:42-55 (2019). PubMed: 30379607