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Epigenetics and Nuclear Signaling Histones H3 Methylated
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Anti-Histone H3 (di methyl K4, tri methyl K4) antibody [mAbcam 6000] - ChIP Grade (ab6000)

  • Datasheet
Reviews (7)Q&A (1)References (31)

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ChIP - Anti-Histone H3 (di methyl K4, tri methyl K4) antibody [mAbcam 6000] - ChIP Grade (ab6000)
  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K4, tri methyl K4) antibody [mAbcam 6000] - ChIP Grade (ab6000)
  • Western blot - Anti-Histone H3 (di methyl K4, tri methyl K4) antibody [mAbcam 6000] - ChIP Grade (ab6000)
  • Flow Cytometry - Anti-Histone H3 (di methyl K4, tri methyl K4) antibody [mAbcam 6000] - ChIP Grade (ab6000)
  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K4, tri methyl K4) antibody [mAbcam 6000] - ChIP Grade (ab6000)

Key features and details

  • Mouse monoclonal [mAbcam 6000] to Histone H3 (di methyl K4, tri methyl K4) - ChIP Grade
  • Suitable for: ICC/IF, WB, ChIP, Flow Cyt
  • Reacts with: Cow, Human, Rice
  • Isotype: IgG2b

You may also be interested in

Protein
Product image
Recombinant human Histone H3 protein (Active) (ab198757)
Secondary
Product image
Goat Anti-Mouse IgG H&L (HRP) (ab205719)

View more associated products

Overview

  • Product name

    Anti-Histone H3 (di methyl K4, tri methyl K4) antibody [mAbcam 6000] - ChIP Grade
    See all Histone H3 primary antibodies
  • Description

    Mouse monoclonal [mAbcam 6000] to Histone H3 (di methyl K4, tri methyl K4) - ChIP Grade
  • Host species

    Mouse
  • Specificity

    This antibody detects a band of the appropriate size when used in Western blotting on a calf thymus histone preparation (see image). It is specifically blocked by peptides corresponding to di methylated K4 of Histone H3 and to tri methylated K4. Slight cross reactivity is observed with mono methyl K4 peptide in ELISA and Western Blot.

  • Tested Applications & Species

    Application Species
    ChIP
    Human
    Flow Cyt
    Human
    ICC/IF
    Human
    WB
    Cow
    See all applications and species data
  • Immunogen

    Synthetic peptide within Human Histone H3 aa 1-100 (tri methyl K4) conjugated to keyhole limpet haemocyanin (Sulfosuccinimidyl 4-N-maleimidomethyl-cyclohexane-1-carboxylate (Sulfo-SMCC)). The exact sequence is proprietary. Hybridomas were prepared and the resulting clones were positively screened by ELISA against the immunising peptide and negatively screened against the non-modified equivalent peptide.
    (Peptide available as ab1342, ab7768)

  • Positive control

    • ChIP: Chromatin was prepared from U-2 OS cell . Flow Cyt: HeLa cells. WB: Calf thymus histone preparation nuclear lysate.
  • General notes

    This antibody clone is manufactured by Abcam.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

    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 6000
  • Myeloma

    Sp2/0-Ag14
  • Isotype

    IgG2b
  • Light chain type

    kappa
  • Research areas

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

Associated products

  • Alternative Versions

    • Alexa Fluor® 647 Anti-Histone H3 (di methyl K4, tri methyl K4) antibody [mAbcam 6000] (ab204246)
    • Alexa Fluor® 488 Anti-Histone H3 (di methyl K4, tri methyl K4) antibody [mAbcam 6000] (ab205814)
    • Anti-Histone H3 (di methyl K4, tri methyl K4) antibody [mAbcam 6000] - BSA and Azide free (ab237973)
  • 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)
    • Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed (ab96879)
  • Corresponding Unmodified Peptide

    • Human Histone H3 (unmodified ) peptide (ab2903)
  • Immunizing Peptide (Blocking)

    • Human Histone H3 (tri methyl K4) peptide (ab1342)
    • Human Histone H3 (di methyl K4) peptide (ab7768)
  • Isotype control

    • Mouse IgG2b, kappa monoclonal [7E10G10] - Isotype Control (ab170192)
  • Recombinant Protein

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

    • Anti-Histone H3 (di methyl K4) antibody - ChIP Grade (ab7766)
    • Anti-Histone H3 (tri methyl K4) antibody - ChIP Grade (ab8580)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab6000 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
Flow Cyt
Human
ICC/IF
Human
WB
Cow
All applications
Mouse
Rat
Rabbit
Chicken
Saccharomyces cerevisiae
Xenopus laevis
Arabidopsis thaliana
Caenorhabditis elegans
Drosophila melanogaster
Schizosaccharomyces pombe
Neurospora crassa
Candida albicans
Application Abreviews Notes
ICC/IF (4)
Use a concentration of 1 µg/ml.
WB (2)
1/500. Predicted molecular weight: 15.2 kDa.Can be blocked with Human Histone H3 (tri methyl K4) peptide (ab1342) or Human Histone H3 (di methyl K4) peptide (ab7768).
ChIP (1)
Use at an assay dependent concentration.
Flow Cyt
Use 1µg for 106 cells.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

Notes
ICC/IF
Use a concentration of 1 µg/ml.
WB
1/500. Predicted molecular weight: 15.2 kDa.Can be blocked with Human Histone H3 (tri methyl K4) peptide (ab1342) or Human Histone H3 (di methyl K4) peptide (ab7768).
ChIP
Use at an assay dependent concentration.
Flow Cyt
Use 1µg for 106 cells.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

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

  • ChIP - Anti-Histone H3 (di methyl K4, tri methyl K4) antibody [mAbcam 6000] - ChIP Grade (ab6000)
    ChIP - Anti-Histone H3 (di methyl K4, tri methyl K4) antibody [mAbcam 6000] - ChIP Grade (ab6000)

    Chromatin was prepared from U2OS cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10min. The  ChIP was performed with 25 µg of chromatin, 2 µg of  ab6000 (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). Primers and probes are located in the first kb of the transcribed region.

  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K4, tri methyl K4) antibody [mAbcam 6000] - ChIP Grade (ab6000)
    Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K4, tri methyl K4) antibody [mAbcam 6000] - ChIP Grade (ab6000)This image is courtesy of Luke Hughes-Davies and Rhiannon Jade, Gurdon Institute, Cambridge, UK

    Immunofluorescent imaging of human cells (U2OS) with ab6000 reveals a diffuse background nuclear staining corresponding to global dimethylation of K4, with multiple foci of brighter staining.  The complete lack of nucleolar or cytoplasmic staining background confirms the high specificity of this antibody in this application.   

    IF was performed with a standard paraformaldehyde technique (fixed in PBS buffered PFH 4% for 5 minutes, permeabilised with 0.5% triton-PBS for 5 minutes, blocked with 5% milk / 0.2% tween for one hour.  Primary antibody used at 1/200 in 5% milk / 0.2% TWEEN for one hour, secondary antibody Alexa 488 for 30 minutes.  All blocking and incubation steps carried out at 37 degrees.

    Due to the size constraints for images on our website, unfortunately we are unable to show a higher quality image.

  • Western blot - Anti-Histone H3 (di methyl K4, tri methyl K4) antibody [mAbcam 6000] - ChIP Grade (ab6000)
    Western blot - Anti-Histone H3 (di methyl K4, tri methyl K4) antibody [mAbcam 6000] - ChIP Grade (ab6000)
    All lanes : Anti-Histone H3 (di methyl K4, tri methyl K4) antibody [mAbcam 6000] - ChIP Grade (ab6000) at 1 µg/ml

    Lane 1 : Calf Thymus Histone Preparation Nuclear Lysate
    Lane 2 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (unmodified) peptide (ab7228) at 0.5 µg
    Lane 3 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (mono methyl K4) peptide (ab1340) at 0.5 µg
    Lane 4 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (di methyl K4) peptide (ab7768) at 0.5 µg
    Lane 5 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (tri methyl K4) peptide (ab1342) at 0.5 µg
    Lane 6 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (mono methyl K9) peptide (ab1771) at 0.5 µg
    Lane 7 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (di methyl K9) peptide (ab1772) at 0.5 µg
    Lane 8 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (tri methyl K9) peptide (ab1773) at 0.5 µg
    Lane 9 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (mono methyl K27) peptide (ab1780) at 0.5 µg
    Lane 10 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (di methyl K27) peptide (ab1781) at 0.5 µg
    Lane 11 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (tri methyl K27) peptide (ab1782) at 0.5 µg

    Lysates/proteins at 0.5 µg per lane.

    Secondary
    All lanes : Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 15.2 kDa
    Observed band size: 17 kDa
    why is the actual band size different from the predicted?


    Exposure time: 20 minutes
  • Flow Cytometry - Anti-Histone H3 (di methyl K4, tri methyl K4) antibody [mAbcam 6000] - ChIP Grade (ab6000)
    Flow Cytometry - Anti-Histone H3 (di methyl K4, tri methyl K4) antibody [mAbcam 6000] - ChIP Grade (ab6000)

    Overlay histogram showing HeLa cells stained with ab6000 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab6000, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K4, tri methyl K4) antibody [mAbcam 6000] - ChIP Grade (ab6000)
    Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K4, tri methyl K4) antibody [mAbcam 6000] - ChIP Grade (ab6000)

    Left image shows human Hep cell monolayer fixed with 4% formaldehyde, permeabilized using 0.5% Triton X-100, blocked with 5% FCS in PBS (20 min) and incubated with ab6000 for 1 hr (diluted 1/200 in PBS plus 1% FCS). Detection with anti-mouse rhodamin labeled secondary antibody. Unstained areas represent nucleoli and heterochromatin as judged from DAPI overlay of the same cell in the right image.

    This image was kindly supplied as part of the review submitted by Christian Schoefer.

Protocols

  • Mouse on Mouse staining protocol
  • ChIP protocols

Click here to view the general protocols

Datasheets and documents

    • Datasheet
  • References (31)

    Publishing research using ab6000? Please let us know so that we can cite the reference in this datasheet.

    ab6000 has been referenced in 31 publications.

    • Liu J  et al. Dynamic patterns of H3K4me3, H3K27me3, and Nanog during rabbit embryo development. Am J Transl Res 11:430-441 (2019). PubMed: 30787999
    • Kang Y  et al. Effects of Salmonella enteritidis Vaccination on the Expression of Innate Immune Molecules and Histone Modifications in the Follicular Theca of Laying Hens. J Poult Sci 56:298-307 (2019). PubMed: 32055228
    • Gibbons HR  et al. Divergent lncRNA GATA3-AS1 Regulates GATA3 Transcription in T-Helper 2 Cells. Front Immunol 9:2512 (2018). PubMed: 30420860
    • Wheaton K  et al. UbE2E1/UBCH6 Is a Critical in Vivo E2 for the PRC1-catalyzed Ubiquitination of H2A at Lys-119. J Biol Chem 292:2893-2902 (2017). PubMed: 28073915
    • Xiong Y  et al. Effects of high progesterone level on the day of human chorionic gonadotrophin administration in in vitro fertilization cycles on epigenetic modification of endometrium in the peri-implantation period. Fertil Steril 108:269-276.e1 (2017). PubMed: 28778281
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-8 of 8 Abreviews or Q&A

    Western blot abreview for Anti-Histone H3 (methylated) antibody [mAbcam 6000] - ChIP Grade

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Hela)
    Loading amount
    10 µg
    Specification
    Hela
    Gel Running Conditions
    Reduced Denaturing (12%)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Mr. Benny Garfinkel

    Verified customer

    Submitted Aug 05 2010

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Histone H3 (methylated) antibody [mAbcam 6000] - ChIP Grade

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (3T3 mouse fibroblasts)
    Specification
    3T3 mouse fibroblasts
    Fixative
    Paraformaldehyde
    Permeabilization
    Yes - 1% Triton X-100 4 minutes
    Read More

    Mr. Benny Garfinkel

    Verified customer

    Submitted Jul 16 2010

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Histone H3 (methylated) antibody [mAbcam 6000] - ChIP Grade

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (Hela)
    Specification
    Hela
    Fixative
    Paraformaldehyde
    Read More

    Mr. Benny Garfinkel

    Verified customer

    Submitted Jul 15 2010

    ChIP abreview for Anti-Histone H3 (methylated) antibody [mAbcam 6000] - ChIP Grade

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    ChIP
    Sample
    Rice Cell lysate - whole cell (14 days old seedlings grown under dark condition)
    Specification
    14 days old seedlings grown under dark condition
    Type
    Native ChIP (N-ChIP)
    Detection step
    Real-time PCR
    Positive control
    Gene of interest
    Negative control
    No Antibody control
    Read More

    豊 佐藤

    Verified customer

    Submitted Dec 05 2007

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Histone H3 (methylated) antibody [mAbcam 6000] - ChIP Grade

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (MEFs SV40 transformed)
    Specification
    MEFs SV40 transformed
    Fixative
    Paraformaldehyde
    Permeabilization
    Yes - 0.5% triton 10 min RT
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: RT°C
    Read More

    Mr. Mark Rochman

    Verified customer

    Submitted Aug 13 2007

    Western blot abreview for Anti-Histone H3 (methylated) antibody [mAbcam 6000] - ChIP Grade

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Not applicable Cell lysate - whole cell (whole cell lysate - Candida albicans)
    Loading amount
    2 µg
    Specification
    whole cell lysate - Candida albicans
    Blocking step
    Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5%
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    Dr. Suresh Babu Raman

    Verified customer

    Submitted Mar 24 2006

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Histone H3 (methylated) antibody [mAbcam 6000] - ChIP Grade

    Inconclusive
    Abreviews
    Abreviews
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (spermatocytes)
    Specification
    spermatocytes
    Fixative
    Paraformaldehyde
    Blocking step
    Other as blocking agent for 30 minute(s) · Concentration: 0.15%BSA+0
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    Abcam user community

    Verified customer

    Submitted Feb 27 2006

    Question

    I'm interesting to purchase antibodies against Histone 3 di methyl K4 (ab7766) or Histone H3 (di+tri methyl K4) ab6000, for the use in ChiP assay. The organism we are working is is Entamoeba histolytica and we know that the sequence around K4 is similar histones from other origins. But we do not know the degree of methylation (di or tri). Would you know any preference of using the di methyl which is polyclonal or the di+tri which is monoclonal for ChIP in order to distinguish between transcriptional active and silenced genes.

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    Abcam community

    Verified customer

    Asked on Nov 21 2005

    Answer

    I have performed an alignment of Entamoeba histolytica and homo sapiens and indeed there does seem to be some similarity in the amino acid sequence at lysine 4 of H3 (ARTK). These antisera have yet to be tested in this species. However this is not to say that it will not work, just that it has yet to be tested. Entamoeba histolytica histone H3 (Q50XG0_ENTHI) Entamoeba MARTKGHIERPSNKSAKAVKNVAFKAA Homo sapi -ARTK-QTARKSTGGKAPRKQLATKAA ****.: * *. . . *::* *** ab7766 and ab6000 are £198 each and have both been tested using our own recommended ChIP protocol. This can be accessed at the following link: https://www.abcam.com/index.html?pageconfig=view_protocol&pid=171 Both antisera work well by ChIP. However, in your example I would recommend determining the presence of either modification using mass spec before embarking on ChIP experiments as negative results are difficult to prove given that these antisera have not been tested on your species of interest.

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    Abcam Scientific Support

    Answered on Nov 22 2005

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