Anti-Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade

• 产品名称

  Anti-Histone H3 (tri methyl K27)抗体[mAbcam 6002] - ChIP Grade
  参阅全部 Histone H3 一抗

• 描述

  小鼠单克隆抗体[mAbcam 6002] to Histone H3 (tri methyl K27) - ChIP Grade

• 宿主

  Mouse

• 特异性

  This antibody is specific for histone H3 tri-methylated at K27. The antibody is blocked in Western blot by tri methyl K27 peptide and slightly by di methyl K27 peptide (there is <12% cross reactivity with di methyl K27 as determined by ELISA). It is not blocked by mono methyl K4, di methyl K4, tri methyl K4, mono methyl K9, di methyl K9, tri methyl K9, mono methyl K27 or unmodified K27 peptides (see the peptide blocking assay blot below).

• 经测试应用

  适用于: ChIP, ELISA, WB, IHC - Wholemount, ICC/IFmore details

• 种属反应性

  与反应: Mouse, Cow, Human, Recombinant fragment
  预测可用于: Rat, Rabbit, Chicken, Xenopus laevis, Arabidopsis thaliana, Drosophila melanogaster, Plants, Zebrafish, Rhesus monkey, Chinese hamster, Rice

• 免疫原

  Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
  (Peptide available as ab1782)

• 阳性对照

• WB: Calf Thymus Histone Preparation Nuclear Lysate, HeLa cell lysates, mouse ES whole cell lysate ChIP: HeLa cells and K562 cells. ICC/IF: HeLa cells. IHC-Wholemount: blastocysts and germ cells and in culture in pluripotent stem cells. ELISA: Histone H3 - tri methyl K27 peptide.

• 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

  mAbcam 6002

• 同种型

  IgG3

• 轻链类型

  kappa

The Abpromise guarantee

Abpromise™承诺保证使用ab6002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

• 

  Western blot - Anti-Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade (ab6002)

  All lanes : Anti-Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade (ab6002) at 1 µg/ml
  
  Lane 1 : Calf Thymus Histone Preparation Nuclear Lysate
  Lane 2 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 peptide (ab17163) at 0.5 µg/ml
  Lane 3 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (mono methyl K4) peptide (ab1340) at 0.5 µg/ml
  Lane 4 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (di methyl K4) peptide (ab7768) at 0.5 µg/ml
  Lane 5 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (tri methyl K4) peptide (ab1342) at 0.5 µg/ml
  Lane 6 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (mono methyl K9) peptide (ab1771) at 0.5 µg/ml
  Lane 7 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (di methyl K9) peptide (ab1772) at 0.5 µg/ml
  Lane 8 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (tri methyl K9) peptide (ab1773) at 0.5 µg/ml
  Lane 9 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (mono methyl K27) peptide (ab1780) at 0.5 µg/ml
  Lane 10 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (di methyl K27) peptide (ab1781) at 0.5 µg/ml
  Lane 11 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (tri methyl K27) peptide (ab1782) at 0.5 µg/ml
  
  Lysates/proteins at 0.25 µg per lane.
  
  Secondary
  All lanes : Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/50000 dilution
  
  Developed using the ECL technique.
  
  Performed under reducing conditions.
  
  Predicted band size: 15 kDa
  Observed band size: 17 kDawhy is the actual band size different from the predicted?
  
  
  Exposure time: 3 minutes
  
  
  

  This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab6002 overnight at 4°C. Antibody binding was detected using an anti-mouse antibody conjugated to HRP, and visualised using ECL development solution ab133406.

• 
  

• 

  IHC - Wholemount - Anti-Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade (ab6002)

  In mice, the X-inactivation process is reversed naturally by X-reactivation in blastocysts and germ cells and in culture in pluripotent stem cells. The image shows late blastocyst-stage mouse embryos consisting of three cell types: epiblast (NANOG-positive, cyan), primitive endoderm (GATA4-positive, red) and trophectoderm (CDX2-positive, green). The inactive X-chromosome (H3K27me3-positive, yellow dots) is reactivated only in the epiblast (cells without yellow spots), which will form the embryo. The germ cell factor PRDM14 and the long noncoding RNA Tsix collaborate during the X-reactivation process in blastocysts and pluripotent stem cells and thereby link epigenetic with cellular reprogramming events.

  Image is courtesy of Bernhard Payer, runner-up of the immunofluorescence imaging competition 2017.

• 

  ELISA - Anti-Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade (ab6002)

  All batches of ab6002 are tested in ELISA against peptides to different Histone H3 modifications. Results show strong binding to Histone H3 - tri methyl K27 peptide (ab1782), indicating that this antibody specifically recognizes the Histone H3 tri methyl K27 modification. Weak binding is also detected against the Histone H3 di methyl K27 modification (<12%) (ab1781).

  ab17163 - Histone H3 - unmodified
  ab1340 - Histone H3 - mono methyl K4
  ab7768 - Histone H3 - di methyl K4
  ab1342 - Histone H3 - tri methyl K4
  ab1771 - Histone H3 - mono methyl K9
  ab1772 - Histone H3 - di methyl K9
  ab1773 - Histone H3 - tri methyl K9
  ab1780 - Histone H3 - mono methyl K27
  ab1781 - Histone H3 - di methyl K27
  ab1782 - Histone H3 - tri methyl K27

• 

  ChIP - Anti-Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade (ab6002)

  Chromatin was prepared from Hela cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 min. The ChIP was performed with 25 µg of chromatin, 5 µg of ab6002 (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 for active and inactive loci, Sybr green approach for heterochromatic loci). Primers and probes are located in the first kb of the transcribed region.
      

• 

  Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade (ab6002)

  Figure showing the nuclear distribution of H3 (tri-methyl K27) antibody, ab6002 in a) a 46 chromosome, XX cell line, and b) a 49 chromosome, XXXXX cell line.

  The location of facultative heterochromatin at the inactive X chromosome is indicated by white arrow heads.

• 

  Western blot - Anti-Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade (ab6002)

  Lanes 1-3 : Anti-Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade (ab6002) at 1 µg/ml (2% BSA)
  Lanes 4-6 : Anti-Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade (ab6002) at 1 µg/ml (3% MILK)
  
  Lanes 1 & 4 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate
  Lanes 2 & 5 : EED-/- mouse ES Whole Cell Lysate
  Lanes 3 & 6 : WT mouse ES Whole Cell Lysate
  
  Lysates/proteins at 10 µg per lane.
  
  Secondary
  All lanes : Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/50000 dilution
  
  Developed using the ECL technique.
  
  Performed under reducing conditions.
  
  Predicted band size: 15 kDa
  Observed band size: 17 kDawhy is the actual band size different from the predicted?
  
  
  Exposure time: 12 minutes
  
  
  

  This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin (lanes 1-3) and 3% milk (lanes 4-6) before being incubated with ab6002 overnight at 4°C. Antibody binding was detected using an anti-mouse antibody conjugated to HRP, and visualised using ECL development solution ab133406.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade (ab6002)

  ICC/IF image of ab6002 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab6002, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) Hek293, HepG2 and MCF7 cells at 5µg/ml, and in 100% methanol fixed (5 min) HeLa, Hek293, HepG2 and MCF7 cells at 5µg/ml.

• 

  ChIP - Anti-Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade (ab6002)This image is courtesy of an anonymous abreview.

  Chromatin was prepared from K562 cells. Cells were fixed with formaldehyde for 10 min. A GATA1 antibody was used as the positive control and a Rabbit IgG was used as the negative control. Incubation with primary antibody was in Immuno Precipitation Dilution buffer for 16 hours at 4°C. The immunoprecipitated DNA was quantified by real time PCR. 

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