Anti-CENPE 抗体 [1H12] (ab5093)
Key features and details
- Mouse monoclonal [1H12] to CENPE
- Suitable for: ICC/IF, WB, Flow Cyt
- Reacts with: Human
- Isotype: IgG1
製品の概要
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製品名
Anti-CENPE antibody [1H12]
CENPE 一次抗体 製品一覧 -
製品の詳細
Mouse monoclonal [1H12] to CENPE -
由来種
Mouse -
アプリケーション
適用あり: ICC/IF, WB, Flow Cytmore details -
種交差性
交差種: Human -
免疫原
Recombinant full length protein (Human).
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ポジティブ・コントロール
- Any human cell line should be suitable as a positive control. Kinetochore staining only visible in mitosis.
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特記事項
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
製品の特性
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製品の状態
Liquid -
保存方法
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. -
バッファー
Preservative: 0.1% Sodium azide
Constituent: PBS -
Concentration information loading...
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特記事項(精製)
Purified from tissue culture supernatant via ion exchange chromatography (>95% total IgG). -
ポリ/モノ
モノクローナル -
クローン名
1H12 -
ミエローマ
Sp2/0 -
アイソタイプ
IgG1 -
軽鎖の種類
kappa -
研究分野
関連製品
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Compatible Secondaries
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Isotype control
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Recombinant Protein
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab5093の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
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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. |
特記事項 |
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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. |
ターゲット情報
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機能
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. -
関連疾患
Microcephaly 13, primary, autosomal recessive -
配列類似性
Belongs to the TRAFAC class myosin-kinesin ATPase superfamily. Kinesin family.
Contains 1 kinesin motor domain. -
ドメイン
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. -
翻訳後修飾
The 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. -
細胞内局在
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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参照データベース
- Entrez Gene: 1062 Human
- Omim: 117143 Human
- SwissProt: Q02224 Human
- Unigene: 75573 Human
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別名
- CENP E antibody
- CENP-E antibody
- CENPE antibody
see all
画像
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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.
プロトコール
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (35)
ab5093 は 35 報の論文で使用されています。
- 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