Anti-Histone H3 (mono methyl K36)抗体- ChIP Grade (ab9048)

概述

  • 产品名称
    Anti-Histone H3 (mono methyl K36)抗体- ChIP Grade
    参阅全部 Histone H3 一抗
  • 描述
    兔多克隆抗体to Histone H3 (mono methyl K36) - ChIP Grade
  • 特异性
    Specific for human Histone H3 mono methyl K36. Shows partial cross-reactivity with di-methyl K36 (please see Western Blot image). This antibody may not be suitable for experiments on yeast lysate. Although the antibody is specifically blocked using the immunising peptide, customer feedback indicates that it detects a band using S. cerevisiae K36 point mutants. We welcome further customer feedback.
  • 经测试应用
    适用于: IHC-P, ICC, ICC/IF, WB, ChIPmore details
  • 种属反应性
    与反应: Mouse, Cow, Human, Pig, Saccharomyces cerevisiae, Arabidopsis thaliana, Drosophila melanogaster
    预测可用于: Mammal
  • 免疫原

    Synthetic peptide within Human Histone H3 aa 1-100 (mono methyl K36) conjugated to Keyhole Limpet Haemocyanin (KLH). The exact sequence is proprietary.
    (Peptide available as ab1783)

  • 阳性对照
    • WB: HeLa histone preparation and HeLa whole cell extract. ChIP: HeLa cells.

性能

  • 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
    Constituent: PBS

    Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
  • Concentration information loading...
  • 纯度
    Immunogen affinity purified
  • 克隆
    多克隆
  • 同种型
    IgG
  • 研究领域

应用

Our Abpromise guarantee covers the use of ab9048 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
IHC-P Use at an assay dependent concentration.
ICC Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration. PubMed: 19727073
WB 1/1000. Can be blocked with Human Histone H3 (mono methyl K36) peptide (ab1783).
ChIP Use 2 µg for 25 µg of chromatin.

靶标

  • 功能
    Core component of nucleosome. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling.
  • 序列相似性
    Belongs to the histone H3 family.
  • 发展阶段
    Expressed during S phase, then expression strongly decreases as cell division slows down during the process of differentiation.
  • 翻译后修饰
    Acetylation is generally linked to gene activation. Acetylation on Lys-10 (H3K9ac) impairs methylation at Arg-9 (H3R8me2s). Acetylation on Lys-19 (H3K18ac) and Lys-24 (H3K24ac) favors methylation at Arg-18 (H3R17me).
    Citrullination at Arg-9 (H3R8ci) and/or Arg-18 (H3R17ci) by PADI4 impairs methylation and represses transcription.
    Asymmetric dimethylation at Arg-18 (H3R17me2a) by CARM1 is linked to gene activation. Symmetric dimethylation at Arg-9 (H3R8me2s) by PRMT5 is linked to gene repression. Asymmetric dimethylation at Arg-3 (H3R2me2a) by PRMT6 is linked to gene repression and is mutually exclusive with H3 Lys-5 methylation (H3K4me2 and H3K4me3). H3R2me2a is present at the 3' of genes regardless of their transcription state and is enriched on inactive promoters, while it is absent on active promoters.
    Methylation at Lys-5 (H3K4me), Lys-37 (H3K36me) and Lys-80 (H3K79me) are linked to gene activation. Methylation at Lys-5 (H3K4me) facilitates subsequent acetylation of H3 and H4. Methylation at Lys-80 (H3K79me) is associated with DNA double-strand break (DSB) responses and is a specific target for TP53BP1. Methylation at Lys-10 (H3K9me) and Lys-28 (H3K27me) are linked to gene repression. Methylation at Lys-10 (H3K9me) is a specific target for HP1 proteins (CBX1, CBX3 and CBX5) and prevents subsequent phosphorylation at Ser-11 (H3S10ph) and acetylation of H3 and H4. Methylation at Lys-5 (H3K4me) and Lys-80 (H3K79me) require preliminary monoubiquitination of H2B at 'Lys-120'. Methylation at Lys-10 (H3K9me) and Lys-28 (H3K27me) are enriched in inactive X chromosome chromatin.
    Phosphorylated at Thr-4 (H3T3ph) by GSG2/haspin during prophase and dephosphorylated during anaphase. Phosphorylation at Ser-11 (H3S10ph) by AURKB is crucial for chromosome condensation and cell-cycle progression during mitosis and meiosis. In addition phosphorylation at Ser-11 (H3S10ph) by RPS6KA4 and RPS6KA5 is important during interphase because it enables the transcription of genes following external stimulation, like mitogens, stress, growth factors or UV irradiation and result in the activation of genes, such as c-fos and c-jun. Phosphorylation at Ser-11 (H3S10ph), which is linked to gene activation, prevents methylation at Lys-10 (H3K9me) but facilitates acetylation of H3 and H4. Phosphorylation at Ser-11 (H3S10ph) by AURKB mediates the dissociation of HP1 proteins (CBX1, CBX3 and CBX5) from heterochromatin. Phosphorylation at Ser-11 (H3S10ph) is also an essential regulatory mechanism for neoplastic cell transformation. Phosphorylated at Ser-29 (H3S28ph) by MLTK isoform 1, RPS6KA5 or AURKB during mitosis or upon ultraviolet B irradiation. Phosphorylation at Thr-7 (H3T6ph) by PRKCBB is a specific tag for epigenetic transcriptional activation that prevents demethylation of Lys-5 (H3K4me) by LSD1/KDM1A. At centromeres, specifically phosphorylated at Thr-12 (H3T11ph) from prophase to early anaphase, by DAPK3 and PKN1. Phosphorylation at Thr-12 (H3T11ph) by PKN1 is a specific tag for epigenetic transcriptional activation that promotes demethylation of Lys-10 (H3K9me) by KDM4C/JMJD2C. Phosphorylation at Tyr-42 (H3Y41ph) by JAK2 promotes exclusion of CBX5 (HP1 alpha) from chromatin.
    Monoubiquitinated by RAG1 in lymphoid cells, monoubiquitination is required for V(D)J recombination (By similarity). Ubiquitinated by the CUL4-DDB-RBX1 complex in response to ultraviolet irradiation. This may weaken the interaction between histones and DNA and facilitate DNA accessibility to repair proteins.
  • 细胞定位
    Nucleus. Chromosome.
  • Information by UniProt
  • 数据库链接
  • 别名
    • HIST1 cluster, H3J antibody
    • Histone gene cluster 1, H3 histone family, member E antibody
    • Histone gene cluster 1, H3G antibody
    • H3 histone family, member J antibody
    • HIST1 cluster, H3E antibody
    • HIST1 cluster, H3I antibody
    • Histone gene cluster 1, H3C antibody
    • FLJ92264 antibody
    • H3 histone family, member A antibody
    • H3 histone family, member B antibody
    • H3 histone family, member C antibody
    • H3 histone family, member D antibody
    • H3 histone family, member F antibody
    • H3 histone family, member H antibody
    • H3 histone family, member I antibody
    • H3 histone family, member K antibody
    • H3 histone family, member L antibody
    • H3 histone family, member T antibody
    • H3 histone, family 3A antibody
    • H3.1 antibody
    • H3.3A antibody
    • H3/a antibody
    • H3/b antibody
    • H3/c antibody
    • H3/d antibody
    • h3/f antibody
    • H3/h antibody
    • H3/i antibody
    • H3/j antibody
    • H3/k antibody
    • H3/l antibody
    • H3/t antibody
    • H31_HUMAN antibody
    • H3F1K antibody
    • H3F3 antibody
    • H3F3A antibody
    • H3FA antibody
    • H3FB antibody
    • H3FC antibody
    • H3FD antibody
    • H3FF antibody
    • H3FH antibody
    • H3FI antibody
    • H3FJ antibody
    • H3FK antibody
    • H3FL antibody
    • HIST1 cluster, H3A antibody
    • HIST1 cluster, H3B antibody
    • HIST1 cluster, H3C antibody
    • HIST1 cluster, H3D antibody
    • HIST1 cluster, H3F antibody
    • HIST1 cluster, H3G antibody
    • HIST1 cluster, H3H 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 1, H3b antibody
    • Histone 1, H3c antibody
    • Histone 1, H3d antibody
    • Histone 1, H3e antibody
    • Histone 1, H3f antibody
    • Histone 1, H3g antibody
    • Histone 1, H3h antibody
    • Histone 1, H3i antibody
    • Histone 1, H3j antibody
    • histone 3, H3 antibody
    • histone cluster 1 H3 family member a antibody
    • histone cluster 1 H3 family member b antibody
    • histone cluster 1 H3 family member c antibody
    • histone cluster 1 H3 family member d antibody
    • histone cluster 1 H3 family member e antibody
    • histone cluster 1 H3 family member f antibody
    • histone cluster 1 H3 family member g antibody
    • histone cluster 1 H3 family member h antibody
    • histone cluster 1 H3 family member i antibody
    • histone cluster 1 H3 family member j antibody
    • Histone cluster 1, H3a antibody
    • Histone cluster 1, H3b antibody
    • Histone cluster 1, H3c antibody
    • Histone cluster 1, H3d antibody
    • Histone cluster 1, H3e antibody
    • Histone cluster 1, H3f antibody
    • Histone cluster 1, H3g antibody
    • Histone cluster 1, H3i antibody
    • Histone cluster 1, H3j antibody
    • Histone gene cluster 1, H3 histone family, member A antibody
    • Histone gene cluster 1, H3 histone family, member B antibody
    • Histone gene cluster 1, H3 histone family, member C antibody
    • Histone gene cluster 1, H3 histone family, member D antibody
    • Histone gene cluster 1, H3 histone family, member F antibody
    • Histone gene cluster 1, H3 histone family, member G antibody
    • Histone gene cluster 1, H3 histone family, member H antibody
    • Histone gene cluster 1, H3 histone family, member I antibody
    • Histone gene cluster 1, H3 histone family, member J antibody
    • Histone gene cluster 1, H3A antibody
    • Histone gene cluster 1, H3B antibody
    • Histone gene cluster 1, H3D antibody
    • Histone gene cluster 1, H3E antibody
    • Histone gene cluster 1, H3F antibody
    • Histone gene cluster 1, H3H antibody
    • Histone gene cluster 1, H3I antibody
    • Histone gene cluster 1, H3J antibody
    • Histone H 3 antibody
    • Histone H3.1 antibody
    • histone H3.1t antibody
    • Histone H3.2 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
    • Histone H3/m antibody
    • Histone H3/o antibody
    see all

图片

  • Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 2µg of ab9048 (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). Primers and probes are located in the first kb of the transcribed region.

  • All lanes : Anti-Histone H3 (mono methyl K36) antibody - ChIP Grade (ab9048) at 1/500 dilution

    Lane 1 : Histone prep
    Lane 2 : Histone prep with Human Histone H3 (mono methyl K36) peptide (ab1783) at 1 µg/ml
    Lane 3 : Histone prep with Human Histone H3 (di methyl K36) peptide (ab1784) at 1 µg/ml
    Lane 4 : Histone prep with Human Histone H3 (tri methyl K36) peptide (ab1785) at 1 µg/ml
    Lane 5 : Histone prep with Human Histone H3 (unmodified ) peptide (ab2623) at 1 µg/ml
    Lane 6 : Histone prep with Human Histone H3 (mono methyl K4) peptide (ab1340) at 1 µg/ml

    Lysates/proteins at 0.5 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/5000 dilution

    Secondary ab: Alexa Fluor 680 Goat anti-rabbit IgG

    0.5µg histone prep used per lane

    Primary antibody:
    Lane 1: ab9048 (Histone H3 Mono Methyl K36) 1/500
    Lane 2: ab9048 (Histone H3 Mono Methyl K36) 1/500 + ab1783 (ab9048) (Histone H3 Mono Methyl K36) peptide 1µg/ml
    Lane 3: ab9048 (Histone H3 Mono Methyl K36) 1/500 + ab1794 (ab9049) (Histone H3 Di Methyl K36) peptide 1µg/ml
    Lane 4: ab9048 (Histone H3 Mono Methyl K36) 1/500 + ab1785 (ab9050) (Histone H3 Tri Methyl K36) peptide 1µg/ml
    Lane 5: ab9048 (Histone H3 Mono Methyl K36) 1/500 + ab2623 (Histone H3 (23-34) – unmodified) peptide 1µg/ml
    Lane 6: ab9048 (Histone H3 Mono Methyl K36) 1/500 + ab1340 (ab8895) (Histone H3 Mono methyl K4) peptide 1µg/ml

    ab9048 specifically recognise

  • ICC/IF image of ab9048 stained HeLa cells. The cells were 4% formaldehyde 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 (ab9048, 0.1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 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% formaldehyde fixed (10 min) Hek293, HepG2 and MCF7 cells at 0.1µg/ml, and in 100% methanol fixed (5 min) HeLa, Hek293, HepG2 and MCF7 cells at 0.1µg/ml.
  • Staining of interphase nuclei of Hela cells with ab9048 (green) at a working dilution of 1/500. The DNA is stained with DAPI. ab9048 appears to be more associated with heterochromatin (DAPI intense regions) than euchromatin (DAPI less intense regions).

  • ab9048 staining Histone H3 (mono methyl K36) in Mouse pancreatic tumor tissue by Immunohistochemistry (Formalin/ PFA-fixed paraffin-embedded tissue sections). The sections were PFA-fixed and subjected to Heat mediated antigen retrieval in citrate and permeabilized with PBST prior to blocking with 5% serum for 1 hour at room temperature. The primary antibody was diluted 1/500 in PBS and incubated with the sample for 8 hours at 4°C. A Biotin-conjugated Goat anti-Rabbit polyclonal was used as the secondary antibody, diluted 1/1000.

    See Abreview

  • All lanes : Anti-Histone H3 (mono methyl K36) antibody - ChIP Grade (ab9048) at 1/1400 dilution

    Lane 1 : Whole cell lysate prepared from Drosophila BG3 cells
    Lane 2 : Whole cell lysate prepared from Drosophila BG3 cells

    Lysates/proteins at 500000 cells per lane.

    Secondary
    HRP donkey anti-rabbit monoclonal at 1/20000 dilution
    Developed using the ECL technique

    Observed band size : 17,42 kDa (why is the actual band size different from the predicted?)


    Exposure time : 30 seconds

    Image courtesy of an anonymous Abreview.

    See Abreview

文献

This product has been referenced in:
  • Janke R  et al. Oncometabolite D-2-Hydroxyglutarate enhances gene silencing through inhibition of specific H3K36 histone demethylases. Elife 6:N/A (2017). WB ; Saccharomyces cerevisiae . Read more (PubMed: 28346139) »
  • Amendola PG  et al. JMJD-5/KDM8 regulates H3K36me2 and is required for late steps of homologous recombination and genome integrity. PLoS Genet 13:e1006632 (2017). WB ; Caenorhabditis elegans . Read more (PubMed: 28207814) »

See all 71 Publications for this product

客户评价及客户问答

Our anti-histone antibodiesare generally tested with 5% BSA in 0.1% TBST for 1 hour at room temperature with gentle rocking when performing western blotting. We also use 5% BSA in the antibody diluentand use with overnight incubation at 4ºC. This proce...

Read More

Each new lot of ab9048 is tested in Western blot with blocking peptides, similarly to the blot shown on the datasheet, and is also tested in IF and ChIP. Results are attached.

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Fruit fly (Drosophila melanogaster) Cell lysate - whole cell (BG3 tissue culture cells)
Loading amount
500000 cells
Specification
BG3 tissue culture cells
Gel Running Conditions
Reduced Denaturing (BioRad TGX 4-20%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Jul 13 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (pancreatic tumor)
Specification
pancreatic tumor
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate
Permeabilization
Yes - PBST
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

提交于 Jan 10 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (primary pancreatic cancer cell line)
Specification
primary pancreatic cancer cell line
Fixative
Paraformaldehyde
Permeabilization
Yes - PBST
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°C
Username

Abcam user community

Verified customer

提交于 Jan 10 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (pancreatic tumor IMPN leasion)
Specification
pancreatic tumor IMPN leasion
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate
Permeabilization
Yes - PBST
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

提交于 Jan 10 2011

Application
Western blot
Sample
Saccharomyces cerevisiae Cell lysate - whole cell (WT and set2 delete S288c S. cerevisiae)
Loading amount
5 µg
Specification
WT and set2 delete S288c S. cerevisiae
Gel Running Conditions
Reduced Denaturing (12.5% SDS-PAGE)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Dr. Marc Meneghini

Verified customer

提交于 Oct 11 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
ChIP
Sample
Human Cell lysate - whole cell (Blood vessel)
Specification
Blood vessel
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: formaldehyde
Detection step
Real-time PCR
Positive control
my samples (GAPDH)
Negative control
my samples
Username

Abcam user community

Verified customer

提交于 Jul 29 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
ChIP
Sample
Saccharomyces cerevisiae Cell lysate - nuclear (yeast)
Specification
yeast
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: CHO
Detection step
Real-time PCR
Positive control
Histone H3 (ab1791)
Negative control
Rabbit IgG
Username

Abcam user community

Verified customer

提交于 Apr 01 2009

Application
Western blot
Sample
Saccharomyces cerevisiae Cell lysate - whole cell (no case)
Loading amount
2 µg
Specification
no case
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

提交于 Oct 23 2007

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