Anti-Dicer antibody [13D6] - ChIP Grade

• 产品名称

  Anti-Dicer抗体[13D6] - ChIP Grade
  参阅全部 Dicer 一抗

• 描述

  小鼠单克隆抗体[13D6] to Dicer - ChIP Grade

• 宿主

  Mouse

• 特异性

  We do not guarantee its use in WB with mouse lysates.

• 经测试应用

  适用于: WB, ChIP, Flow Cytmore details

• 种属反应性

  与反应: Mouse, Human
  不与反应: Caenorhabditis elegans

• 免疫原

  Synthetic peptide corresponding to Human Dicer.

• 阳性对照

• WB: 10ng of recombinant dicer (using ECL or ECL Plus) in under one minute of exposure to film. ICC: Human Fibroblasts

• 常规说明

  To our knowledge the localization of Dicer remains to be fully determined and it appears that its localization can be nuclear and/or cytoplasmic. Please see reference Meltzer, 2005 for a reference to a cytoplasmic localisation (figure 1 of paper), as supported with the staining seen in the image on this datasheet.
  Please note this antibody gave negative results in WB with mouse fibroblast cell line lysates. We do not guarantee its use in WB with mouse lysates.

  
  

  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. Avoid freeze / thaw cycle.

• 存储溶液

  Preservative: 0.02% Sodium azide
  Constituent: HEPES buffered saline

• 浓度

• 100 µg 浓度为 1 mg/ml

• 纯度

  Ammonium Sulphate Precipitation

• 克隆

  单克隆

• 克隆编号

  13D6

• 同种型

  IgG2a

The Abpromise guarantee

Abpromise™承诺保证使用ab14601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: 23405 Human

• Entrez Gene: 192119 Mouse

• Omim: 606241 Human

• SwissProt: Q9UPY3 Human

• SwissProt: Q8R418 Mouse

• Unigene: 738957 Human

• Unigene: 87889 Human

• Unigene: 21135 Mouse

• 别名

• Dicer 1 ribonuclease type III antibody

• Dicer antibody

• DICER_HUMAN antibody

• DICER1 antibody

• Double-strand-specific ribonuclease antibody

• Endoribonuclease Dicer antibody

• Helicase MOI antibody

• Helicase with RNase motif antibody

• HERNA antibody

• KIAA0928 antibody

• MNG1 antibody

• RMSE2 antibody

• DCR antibody

• DCR1 antibody

• Dicer 1 ribonuclease III antibody

• 

  Western blot - Anti-Dicer antibody [13D6] - ChIP Grade (ab14601)

  Lane 1: Wild-type HAP1 cell lysate (20 µg)
  Lane 2: Dicer knockout HAP1 cell lysate (20 µg)

  Lanes 1 and 2: Merged signal (red and green). Green - ab14601 observed at 240 kDa. Red - loading control, ab181602, observed at 37 kDa.

  
  ab14601 was shown to specifically react with Dicer in wild-type HAP1 cells. No band was observed when Dicer knockout samples were used. Wild-type and Dicer knockout samples were subjected to SDS-PAGE. ab14601 and ab181602 (loading control to GAPDH) were diluted 1/1000 and 1/2000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1/10,000 dilution for 1hr at room temperature before imaging.

• 

  Western blot - Anti-Dicer antibody [13D6] - ChIP Grade (ab14601)

  All lanes : Anti-Dicer antibody [13D6] - ChIP Grade (ab14601) at 1/2000 dilution (incubated overnight at 4°C.)
  
  Lane 1 : NIH 3T3
  Lane 2 : A549
  Lane 3 : HepG2
  
  Lysates/proteins at 20 µg per lane.
  
  Secondary
  All lanes : Goat anti-Mouse Green at 1/10000 dilution
  
  
  

  This blot was produced using a 3-8% Tris Acetate gel under the TA buffer system. The gel was run at 150V for 60 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour before being incubated with ab14601 overnight at 4°C. Antibody binding was detected using IR-labelled goat anti-mouse Ab at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.

• 

  Flow Cytometry - Anti-Dicer antibody [13D6] - ChIP Grade (ab14601)

  Overlay histogram showing HEK293 cells stained with ab14601 (red line). The cells were fixed with 80% 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 (ab14601, 0.1μg/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 IgG2a [ICIGG2A] (ab91361, 0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

• 

  Western blot - Anti-Dicer antibody [13D6] - ChIP Grade (ab14601)

  HepG2 cell lysate at 20 µg
  Developed using the ECL technique.
  
  Performed under reducing conditions.
  
  Additional bands at: 100 kDa (possible non-specific binding), 60 kDa (possible non-specific binding)
  
  

• 

  ChIP - Anti-Dicer antibody [13D6] - ChIP Grade (ab14601)This image is courtesy of Elliot Epner MD PhD, Oregon Health and Science University, Portland

  ChIP analysis of Dicer protein binding at chicken ß-globin regulatory elements. Chromatin from human erythroleukemia (K562) cells containing chicken chromosomes with normal (FRK2) or mutant (HS1-K) ß-globin loci and chicken DT40 cells was immunprecipitated with antibodies to Dicer (ab14601). PCR analysis of immunoprecipitated chromatin was carried out using primers to the chicken ß-globin regulatory elements. Mouse IgG served as control. 10% of input DNA was utilized.

• 

  Western blot - Anti-Dicer antibody [13D6] - ChIP Grade (ab14601)This image is courtesy of Jing Huang and Elliot Epner Oregon Health and Science University

  All lanes : Anti-Dicer antibody [13D6] - ChIP Grade (ab14601) at 2.5 µg/ml
  
  Lane 1 : human B cell lymphoma lysate
  Lane 2 : human B cell lymphoma lysate infected with control shRNA retrovirus
  Lane 3 : human B cell lymphoma lysate infected with Dicer knockdown shRNA virus
  
  Lysates/proteins at 2.5 µg/ml per lane.
  
  Observed band size: 225 kDawhy is the actual band size different from the predicted?