Anti-Histone H3 antibody [mAbcam 10799] - ChIP Grade

• 产品名称

  Anti-Histone H3抗体[mAbcam 10799] - ChIP Grade
  参阅全部 Histone H3 一抗

• 描述

  小鼠单克隆抗体[mAbcam 10799] to Histone H3 - ChIP Grade

• 宿主

  Mouse

• 经测试应用

  适用于: ChIP, WB, IHC-P, IPmore details
  不适用于: ICC/IF

• 种属反应性

  与反应: Mouse, Rat, Cow, Human, Drosophila melanogaster, Recombinant fragment
  预测可用于: Monkey, a wide range of other species, Mammals

• 免疫原

  Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
  (Peptide available as ab12149)

• 阳性对照

• WB: HeLa, Calf Thymus Histone and NIH/3T3, whole cell lysates. PC12 nuclear lysate. IP: HeLa whole cell extract; IHC-P: FFPE human breast adenocarcinoma.

• 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

  IgG fraction

• 克隆

  单克隆

• 克隆编号

  mAbcam 10799

• 骨髓瘤

  Sp2/0-Ag14

• 同种型

  IgG3

• 轻链类型

  kappa

The Abpromise guarantee

Abpromise™承诺保证使用ab10799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。

应用说明

Is unsuitable for ICC/IF.

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: 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 antibody [mAbcam 10799] - ChIP Grade (ab10799)

  All lanes : Anti-Histone H3 antibody [mAbcam 10799] - ChIP Grade (ab10799) at 5 µg/ml
  
  Lane 1 : Histone H2A Recombinant Protein at 0.1 µg/ml
  Lane 2 : Histone H2B Recombinant Protein at 0.1 µg/ml
  Lane 3 : Histone H3.1 Recombinant Protein at 0.1 µg/ml
  Lane 4 : Calf Thymus Histone Preparation Nuclear Lysate at 0.5 µg/ml
  Lane 5 : HeLa nuclear lysate (triton enriched) at 10 µg/ml
  Lane 6 : NIH 3T3 nuclear lysate (triton enriched) at 10 µg/ml
  Lane 7 : PC12 nuclear lysate (triton enriched) at 10 µg/ml
  
  Secondary
  All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/5000 dilution
  
  Performed under reducing conditions.
  
  Predicted band size: 15 kDa
  
  
  Exposure time: 20 seconds
  

• 

  ChIP - Anti-Histone H3 antibody [mAbcam 10799] - ChIP Grade (ab10799)

  Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10min. The ChIP was performed with 25µg of chromatin, 2µg of ab10799 (blue), and 20µl 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.    

• 

  Western blot - Anti-Histone H3 antibody [mAbcam 10799] - ChIP Grade (ab10799)

  All lanes : Anti-Histone H3 antibody [mAbcam 10799] - ChIP Grade (ab10799) at 5 µg/ml
  
  Lane 1 : Histone H1 Recombinant Protein
  Lane 2 : Histone H2A Recombinant Protein
  Lane 3 : Histone H2B Recombinant Protein
  Lane 4 : Histone H3.1 Recombinant Protein
  Lane 5 : Histone H4 Recombinant Protein
  
  Lysates/proteins at 0.1 µg per lane.
  
  Secondary
  All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/5000 dilution
  
  Developed using the ECL technique.
  
  Performed under reducing conditions.
  
  Predicted band size: 15 kDa
  
  
  Exposure time: 4 minutes
  
  
  

  We recommend using 3% milk as the blocking agent in Western Blot.

• 

  Immunoprecipitation - Anti-Histone H3 antibody [mAbcam 10799] - ChIP Grade (ab10799)

  Histone H3 [mAbcam 10799] - ChIP Grade was immunoprecipitated using 0.5mg HeLa whole cell extract, 5µg of Mouse monoclonal to and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).

  The antibody was incubated under agitation with Protein G beads for 10min, HeLa whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.

  Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab10799.

  Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/20,000 dilution.

  Band: 15kDa; [mAbcam 10799] Histone H3 - ChIP Grade.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 antibody [mAbcam 10799] - ChIP Grade (ab10799)

  ab10799 staining human kidney sections by IHC-P using EXPOSE IHC detection kit (ab80436). Formalin fixed paraffin embedded tissue sections were pre-treated using heat mediated antigen retrieval (using a pressure cooker) with sodium citrate buffer (pH6) for 30 mins. The section was incubated with ab10799, 5µg/ml, for 1 hour at room temperature. DAB was used as the chromogen and the section was counterstained with haematoxylin and mounted with DPX.

• 

  Western blot - Anti-Histone H3 antibody [mAbcam 10799] - ChIP Grade (ab10799)

  All lanes : Anti-Histone H3 antibody [mAbcam 10799] - ChIP Grade (ab10799) at 1 µg/ml
  
  Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
  Lane 2 : Calf Thymus Histone preparation nuclear lysate
  Lane 3 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate
  
  Lysates/proteins at 10 µg per lane.
  
  Secondary
  All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
  
  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: 90 seconds
  

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 antibody [mAbcam 10799] - ChIP Grade (ab10799)

  IHC image of Histone H3 staining in human breast carcinoma FFPE section, performed on a 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 20 mins. The section was then incubated with ab10799, 1µg/ml, for 8 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.