Anti-CENPE antibody [1H12]

• 产品名称

  Anti-CENPE抗体[1H12]
  参阅全部 CENPE 一抗

• 描述

  小鼠单克隆抗体[1H12] to CENPE

• 宿主

  Mouse

• 经测试应用

  适用于: ICC/IF, WB, Flow Cytmore details

• 种属反应性

  与反应: Human
  

• 免疫原

  Recombinant full length protein (Human).

• 阳性对照

• Any human cell line should be suitable as a positive control. Kinetochore staining only visible in mitosis.

• 常规说明

  
  

  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

• 形式

  Liquid

• 存放说明

  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.

• 存储溶液

  Preservative: 0.1% Sodium azide
  Constituent: PBS

• 浓度

• 100 µl 浓度为 1 mg/ml

• 纯化说明

  Purified from tissue culture supernatant via ion exchange chromatography (>95% total IgG).

• 克隆

  单克隆

• 克隆编号

  1H12

• 骨髓瘤

  Sp2/0

• 同种型

  IgG1

• 轻链类型

  kappa

The Abpromise guarantee

Abpromise™承诺保证使用ab5093于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

• 数据库链接

• Entrez Gene: 1062 Human

• Omim: 117143 Human

• SwissProt: Q02224 Human

• Unigene: 75573 Human

• 别名

• CENPE_HUMAN antibody

• Centromere associated protein E antibody

• Centromere autoantigen E (312kD) antibody

• Centromere autoantigen E antibody

• Centromere protein E 312kDa antibody

• Centromere protein E antibody

• Centromere-associated protein E antibody

• KIF10 antibody

• Kinesin family member 10 antibody

• Kinesin related protein antibody

• Kinesin related protein CENPE antibody

• Kinesin-related protein CENPE antibody

• PPP1R61 antibody

• Protein phosphatase 1, regulatory subunit 61 antibody

• CENP E antibody

• CENP-E antibody

• CENPE antibody

• 

  Immunocytochemistry/ Immunofluorescence - Anti-CENPE antibody [1H12] (ab5093)

  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.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-CENPE antibody [1H12] (ab5093)This image is courtesy of an Abreview submitted by Dr Beth Sullivan

  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.

  See Abreview

• 

  Immunocytochemistry/ Immunofluorescence - Anti-CENPE antibody [1H12] (ab5093)This image is courtesy of Scott Slattery and Mike Mancini

  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.

• 

  Western blot - Anti-CENPE antibody [1H12] (ab5093)

  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 kDawhy is the actual band size different from the predicted?
  
  
  Exposure time: 20 minutes
  

• 

  Flow Cytometry - Anti-CENPE antibody [1H12] (ab5093)

  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.