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Epigenetics and Nuclear Signaling Histones H3 Methylated
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Anti-Histone H3 (tri methyl K27) antibody [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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High-Sensitivity ChIP Kit (ab185913)
Secondary
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Goat Anti-Mouse IgG H&L (HRP) (ab205719)

View more associated products

Overview

  • Product name

    Anti-Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade
    See all Histone H3 primary antibodies
  • Description

    Mouse monoclonal [mAbcam 6002] to Histone H3 (tri methyl K27) - ChIP Grade
  • Host species

    Mouse
  • Specificity

    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).
  • Tested Applications & Species

    Application Species
    ChIP
    Human
    ELISA
    Recombinant fragment
    ICC/IF
    Human
    WB
    Mouse
    Cow
    Human
    See all applications and species data
  • Immunogen

    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)

  • Positive control

    • 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.
  • General notes

    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)

    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 the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

  • Form

    Liquid
  • Storage instructions

    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.
  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituents: PBS, 6.97% L-Arginine
  • Concentration information loading...
  • Purity

    IgG fraction
  • Clonality

    Monoclonal
  • Clone number

    mAbcam 6002
  • Isotype

    IgG3
  • Light chain type

    kappa
  • Research areas

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

Associated products

  • 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)

Applications

The Abpromise guarantee

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.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
ChIP
Human
ELISA
Recombinant fragment
ICC/IF
Human
WB
Mouse
Cow
Human
All applications
Rat
Rabbit
Chicken
Xenopus laevis
Arabidopsis thaliana
Drosophila melanogaster
Plants
Zebrafish
Rhesus monkey
Chinese hamster
Rice
Application Abreviews Notes
ChIP (19)
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 (17)
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 (17)
Use a concentration of 5 µg/ml.
Notes
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.

Target

  • Function

    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.
  • Sequence similarities

    Belongs to the histone H3 family.
  • Developmental stage

    Expressed during S phase, then expression strongly decreases as cell division slows down during the process of differentiation.
  • Post-translational
    modifications

    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.
  • Cellular localization

    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
  • Database links

    • 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
  • Alternative names

    • 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

Images

  • 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

Protocols

  • Mouse on Mouse staining protocol
  • ChIP protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (663)

Publishing research using ab6002? Please let us know so that we can cite the reference in this datasheet.

ab6002 has been referenced in 663 publications.

  • 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

Customer reviews and Q&As

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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

Answered on 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
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Abcam user community

Verified customer

Submitted 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
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Abcam user community

Verified customer

Submitted 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
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Abcam user community

Verified customer

Submitted 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
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Abcam user community

Verified customer

Submitted 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
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Abcam user community

Verified customer

Submitted 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
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Abcam user community

Verified customer

Submitted 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
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Anish Dattani

Verified customer

Submitted 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
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Abcam user community

Verified customer

Submitted 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

Submitted Aug 14 2017

1-10 of 88 Abreviews or Q&A

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