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Epigenetics and Nuclear Signaling Histones H3 Methylated
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Anti-Histone H3 (di methyl K4) 抗体 - ChIP Grade (ab7766)

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Reviews (23)Q&A (9)References (250)

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Abpromise

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ChIP - Anti-Histone H3 (di methyl K4) antibody - ChIP Grade (ab7766)
  • Peptide Array - Anti-Histone H3 (di methyl K4) antibody - ChIP Grade (ab7766)
  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K4) antibody - ChIP Grade (ab7766)
  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K4) antibody - ChIP Grade (ab7766)
  • Western blot - Anti-Histone H3 (di methyl K4) antibody - ChIP Grade (ab7766)
  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K4) antibody - ChIP Grade (ab7766)
  • Western blot - Anti-Histone H3 (di methyl K4) antibody - ChIP Grade (ab7766)

Key features and details

  • Rabbit polyclonal to Histone H3 (di methyl K4) - ChIP Grade
  • Suitable for: ChIP, WB, PepArr, ICC/IF
  • Reacts with: Mouse, Human, Pig, Saccharomyces cerevisiae, Tetrahymena, Drosophila melanogaster, Schizosaccharomyces pombe, Plasmodium falciparum, Common marmoset
  • Isotype: IgG

こちらの製品もご検討ください

タンパク質
Product image
Recombinant human Histone H3 protein (Active) (ab198757)
二次抗体
Product image
Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
一次抗体
Product image
Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - BSA and Azide free (ab222481)

関連製品

製品の概要

  • 製品名

    Anti-Histone H3 (di methyl K4) antibody - ChIP Grade
    Histone H3 一次抗体 製品一覧
  • 製品の詳細

    Rabbit polyclonal to Histone H3 (di methyl K4) - ChIP Grade
  • 由来種

    Rabbit
  • アプリケーション

    適用あり: ChIP, WB, PepArr, ICC/IFmore details
  • 種交差性

    交差種: Mouse, Human, Pig, Saccharomyces cerevisiae, Tetrahymena, Drosophila melanogaster, Schizosaccharomyces pombe, Plasmodium falciparum, Common marmoset
    交差が予測される動物種: Mammals, Candida albicans
  • 免疫原

    Synthetic peptide within Human Histone H3 aa 1-100 (di methyl K4) conjugated to keyhole limpet haemocyanin. The exact sequence is proprietary.
    (Peptide available as ab125304, ab7768)

  • 特記事項

    For detection of Histone H3 specifically methylated at position Lys 4. This antibody was used in a screen by Dover et al, (2002) to isolate yeast mutants that are unable to methylate Lysine 4. In immunofluorescence, this antibody detects foci in the nucleus that are non-colocalising with condensed chromatin. The perinuclear and perinucleolar heterochromatin are not stained with this antibody.

    Learn about ChIP assay kits, other ChIP antibodies, protocols and more in the ChIP assay guide.

    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.

製品の特性

  • 製品の状態

    Liquid
  • 保存方法

    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.
  • バッファー

    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...
  • 精製度

    Immunogen affinity purified
  • 一次抗体 備考

    For detection of Histone H3 specifically methylated at position Lys 4. This antibody was used in a screen by Dover et al, (2002) to isolate yeast mutants that are unable to methylate Lysine 4. In immunofluorescence, this antibody detects foci in the nucleus that are non-colocalising with condensed chromatin. The perinuclear and perinucleolar heterochromatin are not stained with this antibody.
  • ポリ/モノ

    ポリクローナル
  • アイソタイプ

    IgG
  • 研究分野

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

関連製品

  • ChIP Related Products

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

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Immunizing Peptide (Blocking)

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

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Multiplex Immunoassays

    • ALDOA positive control ChIP primer pair (ab269260)
  • Recombinant Protein

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

    • Histone H3 (K4) Methyltransferase Activity Quantification Assay Kit (ab113452)
    • Histone Demethylase (H3K4) Activity Quantification Assay Kit (ab113455)
    • Histone H3 (di-methyl K4) Quantification Kit (Colorimetric) (ab115052)
    • Prestained Protein Ladder - Broad molecular weight (10-245 kDa) (ab116028)
    • Tranylcypromine hydrochloride (2-PCPA), Irreversible monoamine oxidase (MAO) inhibitor (ab120606)

アプリケーション

Our Abpromise guarantee covers the use of ab7766 in the following tested applications.

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

アプリケーション Abreviews 特記事項
ChIP Use 2 µg for 25 µg of chromatin.

Use ALDOA ChIP primer pair ab269260 as positive control.

WB Use a concentration of 1 µg/ml. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa).Can be blocked with Human Histone H3 (di methyl K4) peptide (ab7768).
PepArr Use a concentration of 0.2 - 2 µg/ml.
ICC/IF Use a concentration of 0.1 - 1 µg/ml.

ターゲット情報

  • 機能

    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.
  • 配列類似性

    Belongs to the histone H3 family.
  • 発生段階

    Expressed during S phase, then expression strongly decreases as cell division slows down during the process of differentiation.
  • 翻訳後修飾

    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.
  • 細胞内局在

    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
  • 参照データベース

    • 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
    • Omim: 602810 Human
    • SwissProt: P68431 Human
    • SwissProt: P68433 Mouse
    • 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
    see all
  • 別名

    • 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

画像

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

    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 ab7766 (blue), and 20µl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci). Primers and probes are located in the first kb of the transcribed region.

  • Peptide Array - Anti-Histone H3 (di methyl K4) antibody - ChIP Grade (ab7766)
    Peptide Array - Anti-Histone H3 (di methyl K4) antibody - ChIP Grade (ab7766)

    All batches of ab7766 are tested in Peptide Array against peptides to different Histone H3 modifications. Six dilutions of each peptide are printed on to the Peptide Array in triplicate and results are averaged before being plotted on to a graph. Results show strong binding to Histone H3 - di methyl K4 peptide (ab7768), indicating that this antibody specifically recognises the Histone H3 - di methyl K4 modification.

    1. ab1340 - Histone H3 - mono methyl K4
    2. ab1342 - Histone H3 - tri methyl K4
    3. ab1771 - Histone H3 - mono methyl K9
    4. ab1772 - Histone H3 - di methyl K9
    5. ab1773 - Histone H3 - tri methyl K9
    6. ab1780 - Histone H3 - mono methyl K27
    7. ab1781 - Histone H3 - di methyl K27
    8. ab1782 - Histone H3 - tri methyl K27
    9. ab7228 - Histone H3 - unmodified
    10. ab7768 - Histone H3 - di methyl K4
  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K4) antibody - ChIP Grade (ab7766)
    Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K4) antibody - ChIP Grade (ab7766)

    ab7766 stained in HeLa cells. Cells were fixed with 100% methanol (5 min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% triton for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab7766 at 0.2 µg/ml and ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control) at 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150120 (pseudo-colored red) and ab150081 (colored green) used at 1 µg/ml for 1 hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43 µM for 1 hour at room temperature.

  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K4) antibody - ChIP Grade (ab7766)
    Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K4) antibody - ChIP Grade (ab7766)This image is courtesy of Kirk McManus in the lab of Michael Hendzel, Univeristy of Alberta
    Dimethylated lysine 4 (green) is found in several hundred small nuclear foci that do not colocalize with condensed regions of chromatin (DAPI stained, red).  The perinuclear and perinucleolar heterochromatin do not stain with this antibody.
  • Western blot - Anti-Histone H3 (di methyl K4) antibody - ChIP Grade (ab7766)
    Western blot - Anti-Histone H3 (di methyl K4) antibody - ChIP Grade (ab7766)
    Anti-Histone H3 (di methyl K4) antibody - ChIP Grade (ab7766) at 1 µg/ml + Calf Thymus Histone Preparation Nuclear Lysate (ab121) at 0.5 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 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?
    Additional bands at: 14 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 4 minutes
  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K4) antibody - ChIP Grade (ab7766)
    Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K4) antibody - ChIP Grade (ab7766)This image is courtesy of an Abreview submitted by Dr Alexander Rapp

    ab7766 at 1/200 staining human U2OS (osteosarcoma) cells by ICC/IF. The cells were paraformaldehyde fixed and then stained with the antibody for 1 hour. A Cy2 ® conjugated donkey anti-rabbit antibody was used as the secondary (green). The image shows uniformal staining of the whole nucleus, with several specles found. The insert shows H3-di methyl K4 of tw2 cells only. DAPI nuclear staining is shown in blue.

    See Abreview

  • Western blot - Anti-Histone H3 (di methyl K4) antibody - ChIP Grade (ab7766)
    Western blot - Anti-Histone H3 (di methyl K4) antibody - ChIP Grade (ab7766)

    MCF7 cells were incubated at 37&degC for 24h with vehicle control (0 &microM) and different concentrations of tranylcypromine hydrochloride (ab120606). Increased expression of Histone 3 K4 di-methyl (ab7766) in MCF7 cells correlates with an increase in tranylcypromine hydrochloride concentration, as described in literature.

    Nuclear extracts were prepared with RIPA buffer (containing protease inhibitors and sodium orthovanadate), 10&microg of each were loaded on the gel and the WB was run under reducing conditions. After transfer the membrane was blocked for an hour using 5% BSA before being incubated with ab7766 at 1 &microg /ml and ab1791 at 1 &microg /ml overnight at 4°C. Antibody binding was detected using an anti-rabbit HRP secondary antibody (ab97051) at 1/10000 dilution and visualised using ECL development solution.

     

プロトコール

  • Chromatin Immunofluorescence
  • PCR primers used to test ab7766 in ChIP at Abcam

Click here to view the general protocols

データシートおよび資料

    • Datasheet
  • 参考文献 (250)

    ab7766 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

    ab7766 は 250 報の論文で使用されています。

    • Chong SY  et al. H3K4 methylation at active genes mitigates transcription-replication conflicts during replication stress. Nat Commun 11:809 (2020). PubMed: 32041946
    • Sun H  et al. Metabolic switch and epithelial-mesenchymal transition cooperate to regulate pluripotency. EMBO J 39:e102961 (2020). PubMed: 32090361
    • Song H  et al. Isoform-Specific Lysine Methylation of RORa2 by SETD7 Is Required for Association of the TIP60 Coactivator Complex in Prostate Cancer Progression. Int J Mol Sci 21:N/A (2020). PubMed: 32120841
    • Kumari D  et al. Small Molecules Targeting H3K9 Methylation Prevent Silencing of Reactivated FMR1 Alleles in Fragile X Syndrome Patient Derived Cells. Genes (Basel) 11:N/A (2020). PubMed: 32230785
    • Kwon M  et al. H2B ubiquitylation enhances H3K4 methylation activities of human KMT2 family complexes. Nucleic Acids Res 48:5442-5456 (2020). PubMed: 32365172
    View all Publications for this product

    レビューと Q&A

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    レビューを送る 質問を送る

    1-10 of 32 Abreviews or Q&A

    Western blot abreview for Anti-Histone H3 (di methyl K4) antibody - ChIP Grade

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Pig Tissue lysate - whole (Liver)
    Gel Running Conditions
    Reduced Denaturing (17,5)
    Loading amount
    20 µg
    Specification
    Liver
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 22°C
    Read More

    Abcam user community

    Verified customer

    投稿 Apr 09 2019

    Western blot abreview for Anti-Histone H3 (di methyl K4) antibody - ChIP Grade

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - nuclear (MCF-7 nuclear and cytosolic lysates)
    Gel Running Conditions
    Reduced Denaturing (15% BIS-TRIS gel)
    Loading amount
    10 µg
    Specification
    MCF-7 nuclear and cytosolic lysates
    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

    投稿 May 09 2018

    ChIP abreview for Anti-Histone H3 (di methyl K4) antibody - ChIP Grade

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    ChIP
    Sample
    Human Cell lysate - whole cell (HeLa cells treated with IFNg)
    Negative control
    An adjacent coding region from SMOX genes. Exons of actively transcribed genes are enriched in H3K4me3.
    Specification
    HeLa cells treated with IFNg
    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: 1% formaldehyde on rocker
    Positive control
    Primers for a predicted enhancer in Chr 20 (Cell Cycle,Volume 14, Issue 2, pg253-260 2015). Enhancers have higher deposition of mono-methyl and di-methyl H3K4
    Read More

    Dr. Yupen Chung

    Verified customer

    投稿 Feb 10 2016

    Western blot abreview for Anti-Histone H3 (di methyl K4) antibody - ChIP Grade

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - nuclear (HeLa and HEK 293T nuclear lysate)
    Gel Running Conditions
    Reduced Denaturing (15% SDS gel)
    Loading amount
    10 µg
    Specification
    HeLa and HEK 293T nuclear lysate
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Dr. Yupen Chung

    Verified customer

    投稿 Jan 08 2016

    Western blot abreview for Anti-Histone H3 (di methyl K4) antibody - ChIP Grade

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - nuclear (U87MG Glioblastome cell line)
    Gel Running Conditions
    Reduced Denaturing (15%)
    Loading amount
    1 µg
    Specification
    U87MG Glioblastome cell line
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Read More

    Dr. Martin Engel

    Verified customer

    投稿 Nov 02 2015

    ChIP abreview for Anti-Histone H3 (di methyl K4) antibody - ChIP Grade

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    ChIP
    Detection step
    Real-time PCR
    Sample
    Zebrafish Cell lysate - nuclear (whole embryo shield stage, 6 hours p.f.)
    Specification
    whole embryo shield stage, 6 hours p.f.
    Negative control
    We used PCR for zgc15 and igsf genomic regions (see additional information for primer sequences) as a reference control. No H3K4 mono- and di- methylation was expected in these regions based on previously published microarray data
    Type
    Cross-linking (X-ChIP)
    Duration of cross-linking step: 10 minute(s) and 0 second(s)
    Specification of the cross-linking agent: Formaldehyde 1% in PBST
    Positive control
    ChIP sample was 22 times enriched over input control in hoxb3a and 8 times in tiparp genomic region (see additional information for primer sequences). 10-20x enrichment in H3K4 mono- and di- methylation was expected in these regions based on previously published microarray data.
    Read More

    Mrs. Daria Onichtchouk

    Verified customer

    投稿 Apr 01 2015

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Histone H3 (di methyl K4) antibody - ChIP Grade

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Blocking step
    BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 25°C
    Sample
    Human Cell (HeLa)
    Specification
    HeLa
    Permeabilization
    Yes - tween-20
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    投稿 Jan 12 2015

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Histone H3 (di methyl K4) antibody - ChIP Grade

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step
    BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 25°C
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: pH 6.0 citric acid
    Sample
    Human Tissue sections (Human invasive breast carcinoma)
    Specification
    Human invasive breast carcinoma
    Permeabilization
    Yes - tween-20
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    投稿 Jan 09 2015

    Western blot abreview for Anti-Histone H3 (di methyl K4) antibody - ChIP Grade

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    15 µg
    Gel Running Conditions
    Reduced Denaturing (12% SDS-PAGE)
    Sample
    Mouse Cell lysate - whole cell (MEL)
    Specification
    MEL
    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

    投稿 Jan 08 2015

    Western blot abreview for Anti-Histone H3 (di methyl K4) antibody - ChIP Grade

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    15 µg
    Gel Running Conditions
    Reduced Denaturing
    Sample
    Human Cell lysate - nuclear (U2OS)
    Specification
    U2OS
    Blocking step
    Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
    Read More

    Abcam user community

    Verified customer

    投稿 May 15 2014

    1-10 of 32 Abreviews or Q&A

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