Anti-Histone H3 (di methyl K4, tri methyl K4) antibody [mAbcam 6000] - ChIP Grade

• 产品名称

  Anti-Histone H3 (di methyl K4，tri methyl K4)抗体[mAbcam 6000] - ChIP Grade
  参阅全部 Histone H3 一抗

• 描述

  小鼠单克隆抗体[mAbcam 6000] to Histone H3 (di methyl K4，tri methyl K4) - ChIP Grade

• 宿主

  Mouse

• 特异性

  This antibody detects a band of the appropriate size when used in Western blotting on a calf thymus histone preparation (see image). It is specifically blocked by peptides corresponding to di methylated K4 of Histone H3 and to tri methylated K4. Slight cross reactivity is observed with mono methyl K4 peptide in ELISA and Western Blot.

• 经测试应用

  适用于: ICC/IF, WB, ChIP, Flow Cyt (Intra)more details

• 种属反应性

  与反应: Cow, Human, Rice
  预测可用于: Mouse, Rat, Rabbit, Chicken, Saccharomyces cerevisiae, Xenopus laevis, Arabidopsis thaliana, Caenorhabditis elegans, Drosophila melanogaster, Schizosaccharomyces pombe, Neurospora crassa, Candida albicans

• 免疫原

  Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
  (Peptide available as ab1342, ab7768)

• 阳性对照

• ChIP: Chromatin was prepared from U-2 OS cell. Flow Cyt (Intra): HeLa cells. WB: Calf thymus histone preparation nuclear lysate.

• 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

  Affinity purified

• 克隆

  单克隆

• 克隆编号

  mAbcam 6000

• 骨髓瘤

  Sp2/0-Ag14

• 同种型

  IgG2b

• 轻链类型

  kappa

The Abpromise guarantee

Abpromise™承诺保证使用ab6000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: 8350 Human

• Entrez Gene: 8351 Human

• Entrez Gene: 8352 Human

• Entrez Gene: 8353 Human

• Entrez Gene: 8354 Human

• Entrez Gene: 8355 Human

• Entrez Gene: 8356 Human

• Entrez Gene: 8357 Human

• Entrez Gene: 8358 Human

• Entrez Gene: 8968 Human

• Entrez Gene: 319152 Mouse

• Entrez Gene: 319153 Mouse

• Entrez Gene: 360198 Mouse

• Entrez Gene: 97908 Mouse

• Entrez Gene: 100364501 Rat

• Entrez Gene: 100365669 Rat

• Entrez Gene: 291159 Rat

• Entrez Gene: 679994 Rat

• Entrez Gene: 680511 Rat

• Entrez Gene: 682330 Rat

• Omim: 602810 Human

• SwissProt: P84229 Chicken

• SwissProt: P68432 Cow

• SwissProt: P68431 Human

• SwissProt: P68433 Mouse

• SwissProt: Q6LED0 Rat

• Unigene: 132854 Human

• Unigene: 247813 Human

• Unigene: 247814 Human

• Unigene: 248176 Human

• Unigene: 443021 Human

• Unigene: 484990 Human

• Unigene: 532144 Human

• Unigene: 533292 Human

• Unigene: 546315 Human

• Unigene: 586261 Human

• Unigene: 591778 Human

• Unigene: 221301 Mouse

• Unigene: 261657 Mouse

• Unigene: 377874 Mouse

• Unigene: 390558 Mouse

• Unigene: 397328 Mouse

• Unigene: 138090 Rat

别名

• H3 histone family member E pseudogene antibody

• H3 histone family, member A antibody

• H3/A antibody

• H31_HUMAN antibody

• H3F3 antibody

• H3FA antibody

• Hist1h3a antibody

• HIST1H3B antibody

• HIST1H3C antibody

• HIST1H3D antibody

• HIST1H3E antibody

• HIST1H3F antibody

• HIST1H3G antibody

• HIST1H3H antibody

• HIST1H3I antibody

• HIST1H3J antibody

• HIST3H3 antibody

• histone 1, H3a antibody

• Histone cluster 1, H3a antibody

• Histone H3 3 pseudogene antibody

• Histone H3.1 antibody

• Histone H3/a antibody

• Histone H3/b antibody

• Histone H3/c antibody

• Histone H3/d antibody

• Histone H3/f antibody

• Histone H3/h antibody

• Histone H3/i antibody

• Histone H3/j antibody

• Histone H3/k antibody

• Histone H3/l antibody

• 

  ChIP - Anti-Histone H3 (di methyl K4, tri methyl K4) antibody [mAbcam 6000] - ChIP Grade (ab6000)

  Chromatin was prepared from U2OS cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10min. The  ChIP was performed with 25 µg of chromatin, 2 µg of  ab6000 (blue), and 20 µl of protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach). Primers and probes are located in the first kb of the transcribed region.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K4, tri methyl K4) antibody [mAbcam 6000] - ChIP Grade (ab6000)This image is courtesy of Luke Hughes-Davies and Rhiannon Jade, Gurdon Institute, Cambridge, UK

  Immunofluorescent imaging of human cells (U2OS) with ab6000 reveals a diffuse background nuclear staining corresponding to global dimethylation of K4, with multiple foci of brighter staining.  The complete lack of nucleolar or cytoplasmic staining background confirms the high specificity of this antibody in this application.   

  IF was performed with a standard paraformaldehyde technique (fixed in PBS buffered PFH 4% for 5 minutes, permeabilised with 0.5% triton-PBS for 5 minutes, blocked with 5% milk / 0.2% tween for one hour.  Primary antibody used at 1/200 in 5% milk / 0.2% TWEEN for one hour, secondary antibody Alexa 488 for 30 minutes.  All blocking and incubation steps carried out at 37 degrees.

  Due to the size constraints for images on our website, unfortunately we are unable to show a higher quality image.

• 

  Western blot - Anti-Histone H3 (di methyl K4, tri methyl K4) antibody [mAbcam 6000] - ChIP Grade (ab6000)

  All lanes : Anti-Histone H3 (di methyl K4, tri methyl K4) antibody [mAbcam 6000] - ChIP Grade (ab6000) at 1 µg/ml
  
  Lane 1 : Calf Thymus Histone Preparation Nuclear Lysate
  Lane 2 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (unmodified) peptide (ab7228) at 0.5 µg
  Lane 3 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (mono methyl K4) peptide (ab1340) at 0.5 µg
  Lane 4 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (di methyl K4) peptide (ab7768) at 0.5 µg
  Lane 5 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (tri methyl K4) peptide (ab1342) at 0.5 µg
  Lane 6 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (mono methyl K9) peptide (ab1771) at 0.5 µg
  Lane 7 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (di methyl K9) peptide (ab1772) at 0.5 µg
  Lane 8 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (tri methyl K9) peptide (ab1773) at 0.5 µg
  Lane 9 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (mono methyl K27) peptide (ab1780) at 0.5 µg
  Lane 10 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (di methyl K27) peptide (ab1781) at 0.5 µg
  Lane 11 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (tri methyl K27) peptide (ab1782) at 0.5 µg
  
  Lysates/proteins at 0.5 µg per lane.
  
  Secondary
  All lanes : Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/10000 dilution
  
  Developed using the ECL technique.
  
  Performed under reducing conditions.
  
  Predicted band size: 15.2 kDa
  Observed band size: 17 kDawhy is the actual band size different from the predicted?
  
  
  Exposure time: 20 minutes
  

• 

  Flow Cytometry (Intracellular) - Anti-Histone H3 (di methyl K4, tri methyl K4) antibody [mAbcam 6000] - ChIP Grade (ab6000)

  Overlay histogram showing HeLa cells stained with ab6000 (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 (ab6000, 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 IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K4, tri methyl K4) antibody [mAbcam 6000] - ChIP Grade (ab6000)

  Left image shows human Hep cell monolayer fixed with 4% formaldehyde, permeabilized using 0.5% Triton X-100, blocked with 5% FCS in PBS (20 min) and incubated with ab6000 for 1 hr (diluted 1/200 in PBS plus 1% FCS). Detection with anti-mouse rhodamin labeled secondary antibody. Unstained areas represent nucleoli and heterochromatin as judged from DAPI overlay of the same cell in the right image.

  This image was kindly supplied as part of the review submitted by Christian Schoefer.