Anti-PLK1 antibody [36-298]

• 产品名称

  Anti-PLK1抗体[36-298]
  参阅全部 PLK1 一抗

• 描述

  小鼠单克隆抗体[36-298] to PLK1

• 宿主

  Mouse

• 经测试应用

  适用于: Flow Cyt (Intra), ICC, WBmore details

• 种属反应性

  与反应: Mouse, Human, Recombinant fragment
  预测可用于: Rat

• 免疫原

  Recombinant full length protein corresponding to Human PLK1. His-PLK1 full length purified from Sf9 cells.
  Database link: P53350

• 表位

  aa330-370.

• 阳性对照

• WB: HEK-293, HeLa S3 or U-2 OS cell lysate ICC: HeLa, HeLa S3, NIH/3T3 or U-2 OS cells Flow Cyt (Intra): HeLa cells.

• 常规说明

  
  

  This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

  
  

  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

  pH: 7.40
  Preservative: 0.02% Sodium azide
  Constituents: PBS, 6.97% L-Arginine

• 浓度

• 100 µg 浓度为 1 mg/ml

• 纯度

  Protein A purified

• 克隆

  单克隆

• 克隆编号

  36-298

• 同种型

  IgG1

The Abpromise guarantee

Abpromise™承诺保证使用ab17057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

  hide

• Unigene: 16525 Mouse

• Unigene: 11034 Rat

• Entrez Gene: 5347 Human

• Entrez Gene: 18817 Mouse

• Entrez Gene: 25515 Rat

• Omim: 602098 Human

• SwissProt: P53350 Human

• SwissProt: Q07832 Mouse

• SwissProt: Q62673 Rat

• Unigene: 592049 Human

• 别名

• PLK-1 antibody

• plk1 antibody

• PLK1_HUMAN antibody

• Polo like kinase 1 antibody

• Polo-like kinase 1 antibody

• Serine/threonine protein kinase 13 antibody

• Serine/threonine protein kinase PLK1 antibody

• Serine/threonine-protein kinase 13 antibody

• Serine/threonine-protein kinase PLK1 antibody

• STPK 13 antibody

• STPK13 antibody

• Cell cycle regulated protein kinase antibody

• PLK 1 antibody

• PLK antibody

• 

  Immunocytochemistry - Anti-PLK1 antibody [36-298] (ab17057)

  ab17057 staining PLK1 in HeLa cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab17057 at 1µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

  Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.

• 

  Western blot - Anti-PLK1 antibody [36-298] (ab17057)

  All lanes : Anti-PLK1 antibody [36-298] (ab17057)
  
  Lane 1 : Recombinant PLK1
  Lane 2 : U-2 OS (Human bone osteosarcoma epithelial cell line) cell extract
  Lane 3 : HeLa S3 cell extract
  
  Performed under reducing conditions.
  
  Predicted band size: 68 kDa
  Observed band size: 66 kDawhy is the actual band size different from the predicted?
  
  
  
  

  10% SDS-PAGE gel.

• 

  Immunocytochemistry - Anti-PLK1 antibody [36-298] (ab17057)

  Immunofluoresence using ab17057 and either HeLa S3, NIH/3T3 (Mouse embyro fibroblast cell line) or U-2 OS (Human bone osteosarcoma epithelial cell line) cells.

• 

  Western blot - Anti-PLK1 antibody [36-298] (ab17057)

  Anti-PLK1 antibody [36-298] (ab17057) at 1 µg/ml + HEK-293 (Human epithelial cell line from embryonic kidney) cell lysate at 20 µg
  
  Secondary
  Rabbit Anti-Mouse IgG H&L (HRP) (ab6728) at 1/5000 dilution
  
  Developed using the ECL technique.
  
  Performed under reducing conditions.
  
  Predicted band size: 68 kDa
  Observed band size: 66 kDawhy is the actual band size different from the predicted?
  
  
  Exposure time: 2 minutes
  

• 

  Immunocytochemistry - Anti-PLK1 antibody [36-298] (ab17057)This image is courtesy of Scott Slattery and Mike Mancini

  In panel one HeLa (Human epithelial cell line from cervix adenocarcinoma) cells were stained with ab17057 (green) and DAPI. In the second panel, cells were stained with ab17057 (green) and SH-CREST (red), which stains the centromeres. Fix 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. Permeabilized for 30 minutes with 0.5% TX-100 in PEM. Block 30 minutes in 5% milk in TBST. Primary antibody incubated overnight at 4°C diluted 1/400 in 5% milk in TBST. Secondary antibody incubated 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.

• 

  Flow Cytometry (Intracellular) - Anti-PLK1 antibody [36-298] (ab17057)

  Flow cytometry overlay histogram showing HeLa cells stained with ab17057 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab17057) (1x106 in 100 µl at 5 µg/ml) for 30 min at 22°C.

  The secondary antibody Goat anti-mouse IgG H&L (Alexa Fluor® 488, pre-adsorbed) (ab150117) was used at 1/4000 dilution for 30 min at 22°C.

  Isotype control antibody (black line) was mouse IgG1 kappa; (ab170190) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

  Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

  This antibody gave a positive signal in HeLa cells fixed with 80% methanol (5 min) / permeabilized with 0.1% PBS-Triton X-100 for 15 min used under the same conditions.