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Epigenetics and Nuclear Signaling Histones H3 Methylated

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

  • Datasheet
  • SDS
Reviews (66)Q&A (22)References (663)

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Western blot - Anti-Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade (ab6002)
  • IHC - Wholemount - Anti-Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade (ab6002)
  • ELISA - Anti-Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade (ab6002)
  • ChIP - Anti-Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade (ab6002)
  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade (ab6002)
  • Western blot - Anti-Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade (ab6002)
  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade (ab6002)
  • ChIP - Anti-Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade (ab6002)

Key features and details

  • Mouse monoclonal [mAbcam 6002] to Histone H3 (tri methyl K27) - ChIP Grade
  • Suitable for: ChIP, ELISA, WB, IHC - Wholemount, ICC/IF
  • Reacts with: Mouse, Cow, Human, Recombinant fragment
  • Isotype: IgG3

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概述

  • 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 within Human Histone H3 aa 1-100 (tri methyl K27) conjugated to keyhole limpet haemocyanin (Sulfosuccinimidyl 4-N-maleimidomethyl-cyclohexane-1-carboxylate (Sulfo-SMCC)). The exact sequence is proprietary. Clones were positively screened by ELISA against the immunising peptide. Clones were negatively screened against both the unmodified peptide and also against tri methyl K9 Histone H3 peptide.
    (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

    Western blot protocol advice:

    For the blocking step, we recommend using 3% milk for 1 hour at room temperature. This step may need to be optimized depending on your experimental conditions.

    This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

    We have the following conjugates available:
    Anti-Histone H3 (tri methyl K27) antibody (Alexa Fluor® 488) [mAbcam 6002] (ab205728)
    Anti-Histone H3 (tri methyl K27) antibody (Alexa Fluor® 647) [mAbcam 6002] (ab205729)

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    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, as well as customer reviews and Q&As.

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
    Constituents: PBS, 6.97% L-Arginine
  • Concentration information loading...
  • 纯度

    IgG fraction
  • 克隆

    单克隆
  • 克隆编号

    mAbcam 6002
  • 同种型

    IgG3
  • 轻链类型

    kappa
  • 研究领域

    • Epigenetics and Nuclear Signaling
    • Histones
    • H3
    • Methylated
    • Epigenetics and Nuclear Signaling
    • ChIP assays
    • ChIP antibodies

相关产品

  • ChIP Related Products

    • Rabbit Anti-Mouse IgG H&L (ab46540)
    • ChIP Kit (ab500)
  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
  • Corresponding Unmodified Peptide

    • Human Histone H3 (unmodified ) peptide (ab2623)
  • Immunizing Peptide (Blocking)

    • Human Histone H3 (tri methyl K27) peptide (ab1782)
  • Isotype control

    • Mouse IgG3, kappa monoclonal [MG3-35] - Isotype Control (ab18394)
  • Positive Controls

    • Myo-D positive control ChIP primer pair (ab269261)
  • Recombinant Protein

    • Recombinant human Histone H3 protein (Active) (ab198757)
  • Related Products

    • Histone H3 (K27) Methyltransferase Activity Quantification Assay Kit (ab113454)
    • Histone H3 (tri-methyl K27) Quantification Kit (Colorimetric) (ab115072)
    • Prestained Protein Ladder - Broad molecular weight (10-245 kDa) (ab116028)
    • Human Histone H3 (tri methyl K4) peptide (ab1342)
    • Human Histone H3 (tri methyl K9) peptide (ab1773)
    • Human Histone H3 (unmodified) peptide (ab7228)
    • Human Histone H3 (di methyl K4) peptide (ab7768)

应用

Our Abpromise guarantee covers the use of ab6002 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
ChIP Use 5-10 µg for 25 µg of chromatin.

Use Myo-D ChIP primer pair ab269261 as positive control.

ELISA Use a concentration of 0.025 - 1 µg/ml.
WB Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa).Can be blocked with Human Histone H3 (tri methyl K27) peptide (ab1782).

Blocking: we recommend using 3% milk block for 1 hour at room temperature. This step may need to be optimized for your experiments.

IHC - Wholemount Use at an assay dependent concentration.
ICC/IF Use a concentration of 5 µg/ml.

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.
  • Target information above from: UniProt accession P68431 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 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
    see all
  • 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
    see all

图片

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

    See Abreview

实验方案

  • Mouse on Mouse staining protocol
  • ChIP protocols

Click here to view the general protocols

数据表及文件

    • Datasheet
    • SDS
  • 文献 (663)

    发表研究结果有使用 ab6002?请让我们知道,以便我们可以引用本数据表中的参考文章。

    ab6002 被引用在 663 文献中.

    • Di Giorgio E  et al. Different class IIa HDACs repressive complexes regulate specific epigenetic responses related to cell survival in leiomyosarcoma cells. Nucleic Acids Res 48:646-664 (2020). PubMed: 31754707
    • Karagianni P  et al. Bookmarking by Non-pioneer Transcription Factors during Liver Development Establishes Competence for Future Gene Activation. Cell Rep 30:1319-1328.e6 (2020). PubMed: 32023452
    • Song H  et al. HNF4A-AS1/hnRNPU/CTCF axis as a therapeutic target for aerobic glycolysis and neuroblastoma progression. J Hematol Oncol 13:24 (2020). PubMed: 32216806
    • Lobo-Silva J  et al. The antischistosomal potential of GSK-J4, an H3K27 demethylase inhibitor: insights from molecular modeling, transcriptomics and in vitro assays. Parasit Vectors 13:140 (2020). PubMed: 32178714
    • Wang Z  et al. Modulation of SRSF2 expression reverses the exhaustion of TILs via the epigenetic regulation of immune checkpoint molecules. Cell Mol Life Sci 77:3441-3452 (2020). PubMed: 31838573
    View all Publications for this product

    客户评价及客户问答

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    提交评价 提交问题

    1-10 of 88 Abreviews or Q&A

    Question

    Have you characterized the identity of the second band? I am detecting two bands in my WB as well.

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    Abcam community

    Verified customer

    Asked on Jun 25 2012

    Answer

    I just heard back from that lab that although we haven't confirmed it experimentally, the lower band has always been assumed to be either clipped H3 or weak cross-reaction with H4. So long as it is weak in comparison to the H3 band and all other applications look good we are happy to pass this.

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    Abcam Scientific Support

    回复于 Jun 25 2012

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Lizard Tissue sections (Gonad)
    Antigen retrieval step
    Heat mediated
    Permeabilization
    No
    Specification
    Gonad
    Blocking step
    Opal Blocking Buffer as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    提交于 Jun 13 2019

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

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - other (mouse liver purified histone, HCT116 extracted his)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    1 µg
    Specification
    mouse liver purified histone, HCT116 extracted his
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
    Read More

    Abcam user community

    Verified customer

    提交于 Sep 06 2018

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

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (B Cells)
    Gel Running Conditions
    Reduced Denaturing (4-20%)
    Loading amount
    30 µg
    Specification
    B Cells
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    提交于 Jun 29 2018

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

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    ChIP
    Sample
    Human Cell lysate - nuclear (293T)
    Negative control
    Control IgG
    Specification
    293T
    Detection step
    Real-time PCR
    Type
    Cross-linking (X-ChIP)
    Duration of cross-linking step: 15 minute(s) and 0 second(s)
    Specification of the cross-linking agent: PFA
    Positive control
    GAPDH
    Read More

    Abcam user community

    Verified customer

    提交于 Jun 15 2018

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

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (Hela)
    Permeabilization
    Yes - Triton X-100
    Specification
    Hela
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 22°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    提交于 Jun 15 2018

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

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (293T)
    Gel Running Conditions
    Non-reduced Denaturing (16%)
    Loading amount
    5 µg
    Specification
    293T
    Blocking step
    BSA as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
    Read More

    Abcam user community

    Verified customer

    提交于 Jun 14 2018

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

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Schmidtea mediterranea Cell lysate - nuclear (Whole planarian worm)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    30 µg
    Specification
    Whole planarian worm
    Blocking step
    Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Anish Dattani

    Verified customer

    提交于 Feb 08 2018

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

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    ChIP
    Sample
    Human Cell lysate - nuclear (K562 CML cell line)
    Negative control
    Rabbit IgG
    Specification
    K562 CML cell line
    Detection step
    Real-time PCR
    Type
    Cross-linking (X-ChIP)
    Duration of cross-linking step: 10 minute(s) and 0 second(s)
    Specification of the cross-linking agent: Formaldehyde
    Positive control
    GATA1 antibody
    Read More

    Abcam user community

    Verified customer

    提交于 Sep 29 2017

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

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    ChIP
    Sample
    Human Cell lysate - whole cell (esophagus)
    Negative control
    IgG
    Specification
    esophagus
    Detection step
    Real-time PCR
    Type
    Native ChIP (N-ChIP)
    Positive control
    polymerase ii
    Read More

    Abcam user community

    Verified customer

    提交于 Aug 14 2017

    1-10 of 88 Abreviews or Q&A

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