Anti-Ezrin antibody [3C12]

• 产品名称

  Anti-Ezrin抗体[3C12]
  参阅全部 Ezrin 一抗

• 描述

  小鼠单克隆抗体[3C12] to Ezrin

• 宿主

  Mouse

• 经测试应用

  适用于: WB, IHC-Fr, Flow Cyt (Intra), IHC-Pmore details

• 种属反应性

  与反应: Mouse, Human
  预测可用于: Rat, Hamster, Cow, Pig, Kangaroo, Monkey

• 免疫原

  Recombinant fragment corresponding to Human Ezrin aa 350-600.

• 阳性对照

• Lung.

• 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 long term. Avoid freeze / thaw cycle.

• 存储溶液

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

• 浓度

• 100 µg 浓度为 1 mg/ml

• 纯度

  Affinity purified

• 克隆

  单克隆

• 克隆编号

  3C12

• 同种型

  IgG1

• 轻链类型

  kappa

The Abpromise guarantee

Abpromise™承诺保证使用ab4069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: 281574 Cow

• Entrez Gene: 7430 Human

• Entrez Gene: 22350 Mouse

• Entrez Gene: 54319 Rat

• Omim: 123900 Human

• SwissProt: P31976 Cow

• SwissProt: P15311 Human

• SwissProt: P26040 Mouse

• SwissProt: P31977 Rat

• Unigene: 487027 Human

• Unigene: 277812 Mouse

• Unigene: 773 Rat

别名

• Villin 2 ezrin antibody

• CVIL antibody

• CVL antibody

• Cytovillin 2 antibody

• Cytovillin antibody

• DKFZp762H157 antibody

• Epididymis secretory protein Li 105 antibody

• EZR antibody

• EZRI_HUMAN antibody

• Ezrin antibody

• FLJ26216 antibody

• HEL S 105 antibody

• MGC1584 antibody

• p81 antibody

• VIL 2 antibody

• VIL2 antibody

• Villin 2 (ezrin) antibody

• Villin 2 antibody

• Villin-2 antibody

• Villin2 antibody

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ezrin antibody [3C12] (ab4069)

  IHC image of Ezrin staining in a section of formalin-fixed paraffin-embedded normal human lung performed on a Leica BONDTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab4069, 0.025ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

  For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ezrin antibody [3C12] (ab4069)

  IHC image of Ezrin staining in mouse lung formalin fixed paraffin embedded tissue section. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6) for 30mins. The section was incubated with ab4069, 1µg/ml overnight at +4°C. An HRP-conjugated secondary (Ab97040, 1/1000 dilution) was used for 1hr at room temperature. The section was counterstained with haematoxylin and mounted with DPX.

• 

  Immunohistochemistry (Frozen sections) - Anti-Ezrin antibody [3C12] (ab4069)

  IHC image of ab4069 staining in 10% formaldehyde fixed frozen tissue section of human placenta.

  Non-specific protein-protein interactions were blocked using TBS containing 0.025% (v/v) Triton X-100, 0.3M (w/v) glycine and 3% (w/v) BSA for 1 hour at room temperature. The section was then incubated with ab4069 (1μg/ml dilution) in TBS containing 0.025% (v/v) Triton X-100 and 3% (w/v) BSA overnight at +4°C. The section was then incubated with ab150119 (Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 647)) and DAPI for 1 hour at room temperature.

  The DAPI only control (no antibody) inset shows no autofluorescence, demonstrating that any Alexa Fluor® 647 signal is derived directly from bound ab4069. 

  For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antibody concentrations and incubation times.

• 

  Western blot - Anti-Ezrin antibody [3C12] (ab4069)

  All lanes : Anti-Ezrin antibody [3C12] (ab4069) at 1 µg/ml
  
  Lane 1 : Wild-type HAP1 whole cell lysate
  Lane 2 : EZR (Ezrin) knockout HAP1 whole cell lysate
  Lane 3 : HeLa whole cell lysate
  Lane 4 : MEF whole cell lysate
  Lane 5 : NIH3T3 whole cell lysate
  
  Lysates/proteins at 20 µg per lane.
  
  Predicted band size: 69 kDa
  
  
  
  

  Lanes 1 - 5: Merged signal (red and green). Green - ab4069 observed at 81 kDa. Red - loading control, ab181602, observed at 37 kDa.
  
  ab4069 was shown to specifically react with Ezrin in wild-type HAP1 cells as signal was lost in EZR (Ezrin) knockout cells. Wild-type and EZR (Ezrin) knockout samples were subjected to SDS-PAGE. Ab4069 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at 1 μg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

• 

  Western blot - Anti-Ezrin antibody [3C12] (ab4069)This image is courtesy of an anonymous Abreview

  Anti-Ezrin antibody [3C12] (ab4069) at 1/500 dilution + Mouse bEnd.3 whole cell lysate at 20 µg
  
  Secondary
  HRP-conjugated goat anti-mouse IgG polyclonal at 1/10000 dilution
  
  Developed using the ECL technique.
  
  Performed under reducing conditions.
  
  Predicted band size: 69 kDa
  Observed band size: 81 kDawhy is the actual band size different from the predicted?
  
  
  Exposure time: 10 seconds
  

  See Abreview

• 

  Flow Cytometry (Intracellular) - Anti-Ezrin antibody [3C12] (ab4069)

  Overlay histogram showing SH-SY5Y cells stained with ab4069 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab4069, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. 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. This antibody gave a positive signal in SH-SY5Y cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.