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    products/primary-antibodies/histone-h3-di-methyl-k9-antibody-mabcam-1220-chip-grade-ab1220.pdf

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Epigenetics and Nuclear Signaling Histones H3 Methylated
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Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)

  • Datasheet
  • SDS
Reviews (72)Q&A (12)References (786)

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ChIP - Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)
  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)
  • Western blot - Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)
  • ChIP - Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)
  • ChIP - Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)
  • ChIP - Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)
  • Western blot - Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)
  • ELISA - Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)
  • Western blot - Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)
  • Western blot - Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)

Key features and details

  • Mouse monoclonal [mAbcam 1220] to Histone H3 (di methyl K9) - ChIP Grade
  • Suitable for: ICC/IF, WB, ELISA, IHC-P, ChIP
  • Reacts with: Cow, Human, Arabidopsis thaliana, Drosophila melanogaster, Rice
  • Isotype: IgG2a

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 647 HRP

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Anti-Histone H3 (mono methyl K4) antibody [ERP16597] - ChIP Grade – BSA and Azide free (ab239402)

View more associated products

Overview

  • Product name

    Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade
    See all Histone H3 primary antibodies
  • Description

    Mouse monoclonal [mAbcam 1220] to Histone H3 (di methyl K9) - ChIP Grade
  • Host species

    Mouse
  • Specificity

    By peptide ELISA ab1220 recognizes di methyl K9, but not unmodified K9, mono methyl K9, tri methyl K9, di methyl K27, tri methyl K27, mono methyl K4, di methyl K4 or tri methyl K4. By Western blot ab1220 is blocked by di methyl K9, but not by unmodified K9, mono methyl K9, tri methyl K9, di methyl K27, tri methyl K27, mono methyl K4, di methyl K4 or tri methyl K4. This indicates the specificity of ab1220 for di methyl K9 of Histone H3.
  • Tested applications

    Suitable for: ICC/IF, WB, ELISA, IHC-P, ChIPmore details
  • Species reactivity

    Reacts with: Cow, Human, Arabidopsis thaliana, Drosophila melanogaster, Rice
    Predicted to work with: Mouse, Rat, Sheep, Chicken, Saccharomyces cerevisiae, Xenopus laevis, Caenorhabditis elegans, Schizosaccharomyces pombe, Corn, Common marmoset, Other species
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
    (Peptide available as ab1772)

  • Positive control

    • IHC-P: Human kidney tissue; Mouse liver tissue. WB: HeLa whole cell lysate; Calf thymus histone lysate; HEK-293 lysate. ChIP: AFM ChIP primer pair ab269259; Chromatin prepared from U2OS cells. ICC/IF: HeLa cells.
  • General notes

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

    ChIP protocols:
    ChIP protocol for cross-linking ChIP (X-ChIP)
    Native ChIP protocol
    Chromatin preparation from tissues for ChIP
    ChIP troubleshooting
    ChIP tips and tricks guide

    This antibody clone [mAbcam 1220] 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 this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. 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, along with publications, customer reviews and Q&As

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

    Protein G purified
  • Clonality

    Monoclonal
  • Clone number

    mAbcam 1220
  • Isotype

    IgG2a
  • 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® 488 Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] (ab203850)
    • Alexa Fluor® 647 Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] (ab203851)
    • HRP Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] (ab62168)
  • 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 (ab2903)
  • Immunizing Peptide (Blocking)

    • Human Histone H3 (di methyl K9) peptide (ab1772)
  • Isotype control

    • Mouse IgG2a, kappa monoclonal [MG2a-53] - Isotype control (ab18415)
  • Positive Controls

    • AFM positive control ChIP primer pair (ab269259)
  • Recombinant Protein

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

    • Histone H3 (K9) Methyltransferase Activity Quantification Assay Kit (ab113453)
    • Histone H3 (di-methyl K9) Quantification Kit (Colorimetric) (ab115062)
    • 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 (tri methyl K27) peptide (ab1782)
    • Human Histone H3 (unmodified) peptide (ab7228)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab1220 in the following tested applications.

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

Application Abreviews Notes
ICC/IF (16)
Use a concentration of 5 µg/ml.
WB (33)
Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 17 kDa (predicted molecular weight: 17 kDa).Can be blocked with Human Histone H3 (di methyl K9) peptide (ab1772).
ELISA
Use a concentration of 0.00025 - 1 µg/ml.

when testing with immunogen peptide.

IHC-P (7)
Use at an assay dependent concentration.
ChIP (11)
Use 2-4 µg for 25 µg of chromatin.

Use AFM ChIP primer pair ab269259 as positive control.

Notes
ICC/IF
Use a concentration of 5 µg/ml.
WB
Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 17 kDa (predicted molecular weight: 17 kDa).Can be blocked with Human Histone H3 (di methyl K9) peptide (ab1772).
ELISA
Use a concentration of 0.00025 - 1 µg/ml.

when testing with immunogen peptide.

IHC-P
Use at an assay dependent concentration.
ChIP
Use 2-4 µg for 25 µg of chromatin.

Use AFM ChIP primer pair ab269259 as positive control.

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: P84229 Chicken
    • 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 K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)
    ChIP - Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)Image from Alarcon V et al., Sci Rep 13(6), Fig 4. Doi: 10.1038/srep25901. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    Alarcon V et al investigates the effects of pargyline due to inhibition of LSD1. Levels of LSD1 protein were reduced using siRNA (siLSD1). Huh7 (Human hepatocellular carcinoma) cells were treated with and without siLSD1 and incubated for 24 hours before transfection of HBV genome (A). Western blot analysis shows the reduction of LSD1 after treatment. Covalent post-translational modifications on Histone H3 was determined by ChIP using ab1220 as one of the specific antibodies (C-G). HBV cccDNA and cytoplasmic levels were determined by qPCR (H).

  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)
    Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)

    ab1220 staining Histone H3 (di methyl K9) in HeLa (Human cervix adenocarcinoma epithelial cell) cells. The cells were fixed with 4% PFA (10 mins), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab1220 at 5 µg/ml and ab190573, Rabbit monoclonal to alpha Tubulin (Alexa Fluor® 647), at 2 µg/ml (shown in red). The secondary antibody (shown in green) was ab150117, Alexa Fluor® 488 Goat anti-Mouse IgG (H+L) used at a 1/1000 dilution for 1h at room temperature. Nuclear DNA was labelled with DAPI (shown in blue).
    Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
    This product also gave a positive outcome under the same testing conditions in HeLa cells fixed with 100% methanol (5 mins).

  • Western blot - Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)
    Western blot - Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)
    All lanes : Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)

    Lane 1 : Calf thymus histone lysate
    Lane 2 : Calf thymus histone lysate with Histone H3 peptide - unmodified at 1 µg/ml
    Lane 3 : Calf thymus histone lysate with Human Histone H3 (mono methyl K9) peptide (ab1771) at 1 µg/ml
    Lane 4 : Calf thymus histone lysate with Human Histone H3 (di methyl K9) peptide (ab1772) at 1 µg/ml
    Lane 5 : Calf thymus histone lysate with Human Histone H3 (tri methyl K9) peptide (ab1773) at 1 µg/ml
    Lane 6 : Calf thymus histone lysate with Human Histone H3 (di methyl K4) peptide (ab7768) at 1 µg/ml
    Lane 7 : Calf thymus histone lysate with Human Histone H3 (di methyl K27) peptide (ab1781) at 1 µg/ml

    Secondary
    All lanes : Rabbit Anti-Mouse IgG H&L (HRP) (ab6728) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 17 kDa


    Exposure time: 1 minute
  • ChIP - Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)
    ChIP - Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)

    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 25µg of chromatin, 2µg of  ab1220 (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.
     

  • ChIP - Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)
    ChIP - Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)Image from Chen L et al., PLoS One 9(7), Fig 6. Doi: 10.1371/journal.pone.0101040 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    Chen L et al investigates role of Trim 28 (Tripartite motif containing 28) in the epithelial to mesenchymal transition which is implicated in cancer metastasis. ChIP was performed using ab1220. ChIP assay was performed in Trim 28 knockdown A549 cells and control (A). ChIP assay was performed in control and Trim28 expressed in A549 cells (B).

  • ChIP - Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)
    ChIP - Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)Image from Ebata KT et al., Epigenetics Chromatin 10(36), Fig 2. Doi: 10.1186/s13072-017-0143-3 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/.

    ChIP was performed using ab1220. Naïve Mouse ES cells were resuspended in PBS and crosslinked with 1% formaldehyde for 5 minutes at room temperature. Crosslinking was quenched with 125m M glycine. A mouse anti IgG (ab18413) was used as a control. ChIP -seq for H3K9me2 in ES cells treated with and without vitamin C (image a). H3K9me2 signal genome-wide was plotted comparing untreated and vitamin C-treated cells (image b). Fold change for average H3K9me2 signal at gene promoters. Most gene promoters display a reduction in H3K9me2 in vitamin C-treated ES cells (image c). ChIP-qPCR for H3K9me2 in ES cells with or without vitamin C at gene promoters. ChIP for IgG was performed as a negative control (image d). Asterisks represent P < 0.05 by t test

  • Western blot - Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)
    Western blot - Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)This image is courtesy of an anonymous abreview.
    Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220) at 1/1000 dilution + HEK293 at 30 µg

    Secondary
    HRP conjugated polyclonal Sheep anti-Mouse IgG at 1/5000 dilution

    Performed under reducing conditions.

    Predicted band size: 17 kDa



    Blocking buffer: 5% milk

    See Abreview

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)

    ab1220 staining human kidney sections by IHC-P using EXPOSE IHC detection kit (ab80436). Formalin fixed paraffin embedded tissue sections were pre-treated using heat mediated antigen retrieval (using a pressure cooker) with sodium citrate buffer (pH6) for 30 mins. The section was incubated with ab1220, 5µg/ml, for 1 hour at room temperature. DAB was used as the chromogen and the section was counterstained with haematoxylin and mounted with DPX.

  • ELISA - Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)
    ELISA - Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)

    ELISA using ab1220 at varying antibody concentrations.

    The purple line indicates binding to the Human Histone H3 (di methyl K9) peptide ab1772. Binding to the following peptides was not seen:
    Human Histone H3 (unmodified ) peptide (ab2903),
    Human Histone H3 (mono methyl K9) peptide (ab1771),
    Human Histone H3 (tri methyl K9) peptide (ab1773),
    Human Histone H3 (di methyl K27) peptide (ab1781),
    Human Histone H3 (tri methyl K27) peptide (ab1782),
    Human Histone H3 (mono methyl K4) peptide (ab1340),
    Human Histone H3 (di methyl K4) peptide (ab7768),
    Human Histone H3 (tri methyl K4) peptide (ab1342).

    This indicates the specificity of ab1220 for di methyl K9 of Histone H3.

  • Western blot - Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)
    Western blot - Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)
    Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220) at 1 µg/ml + HeLa (Human epitherlial carcinoma cell line) Whole Cell Lysate at 20 µg

    Secondary
    Goat polyclonal to Mouse IgG-H&L- Pre-Adsorbed (HRP) at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 17 kDa
    Observed band size: 17 kDa


    Exposure time: 5 minutes


    Lane 1 : Marker.

    All blocking and antibody incubation steps were done with 5% milk in 20mM Tris-HCL, and 0.1% TWEEN-20.

  • Western blot - Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)
    Western blot - Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)This image is courtesy of an anonymous Abreview
    All lanes : Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220) at 2.5 µg/ml

    Lane 1 : Fruit fly embryo tissue lysate - nuclear at 4.5 µg
    Lane 2 : Fruit fly embryo tissue lysate - nuclear at 1.5 µg
    Lane 3 : Fruit fly embryo tissue lysate - nuclear at 0.5 µg

    Secondary
    All lanes : HRP-conjugated Goat anti-mouse IgG polyclonal at 1/2000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 17 kDa
    Observed band size: 15 kDa why is the actual band size different from the predicted?


    Exposure time: 1 minute

    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 (786)

Publishing research using ab1220? Please let us know so that we can cite the reference in this datasheet.

ab1220 has been referenced in 786 publications.

  • Liu Q  et al. Histone demethylase PHF8 drives neuroendocrine prostate cancer progression by epigenetically upregulating FOXA2. J Pathol 253:106-118 (2021). PubMed: 33009820
  • Leng X  et al. Epigenetic modification-dependent androgen receptor occupancy facilitates the ectopic TSPY1 expression in prostate cancer cells. Cancer Sci 112:691-702 (2021). PubMed: 33185915
  • Yang Y  et al. Modulators of histone demethylase JMJD1C selectively target leukemic stem cells. FEBS Open Bio 11:265-277 (2021). PubMed: 33289299
  • Velásquez ZD  et al. Eimeria bovis infections induce G1 cell cycle arrest and a senescence-like phenotype in endothelial host cells. Parasitology 148:341-353 (2021). PubMed: 33100232
  • Liu J  et al. Extracellular vesicles-encapsulated let-7i shed from bone mesenchymal stem cells suppress lung cancer via KDM3A/DCLK1/FXYD3 axis. J Cell Mol Med 25:1911-1926 (2021). PubMed: 33350586
View all Publications for this product

Customer reviews and Q&As

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1-10 of 84 Abreviews or Q&A

ChIP abreview for Anti-Histone H3 (methylated) antibody [mAbcam 1220] - ChIP Grade

Excellent
Abreviews
Abreviews
Application
ChIP
Sample
Mouse Cell lysate - whole cell (C2C12 mouse myoblast cells)
Specification
C2C12 mouse myoblast cells
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: 0.75% formaldehyde
Detection step
Real-time PCR
Positive control
Control C2C12 cells at day 2 of differentiation
Negative control
Beads only and mouse IgG were the negative controls
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Apr 29 2013

Immunohistochemistry (Frozen sections) abreview for Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (Colon)
Permeabilization
No
Specification
Colon
Blocking step
Serum as blocking agent for 15 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
Acetone
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Mar 09 2023

Western blot abreview for Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Drosophila melanogaster Tissue lysate - whole (larval brain)
Gel Running Conditions
Reduced Denaturing (15%)
Loading amount
5 µg
Specification
larval brain
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

PROF. Laura Ciapponi

Verified customer

Submitted Nov 15 2022

Immunocytochemistry/ Immunofluorescence abreview for Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade

Good
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Drosophila melanogaster Cell (larval brain cells)
Permeabilization
Yes - PBS-Triton 0,1%
Specification
larval brain cells
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 25°C
Fixative
Formaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

PROF. Laura Ciapponi

Verified customer

Submitted Nov 15 2022

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Cell (fetal germline)
Permeabilization
No
Specification
fetal germline
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Apr 01 2022

Western blot abreview for Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (U2OS)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
U2OS
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jul 15 2021

Western blot abreview for Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (Mouse vascular smooth muscle cells)
Gel Running Conditions
Reduced Denaturing
Loading amount
30 µg
Specification
Mouse vascular smooth muscle cells
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jul 15 2021

Immunocytochemistry/ Immunofluorescence abreview for Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Mouse vascular smooth muscle cells)
Permeabilization
Yes - NP40
Specification
Mouse vascular smooth muscle cells
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
Fixative
Formaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jul 02 2021

Immunocytochemistry/ Immunofluorescence abreview for Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (U2OS)
Permeabilization
Yes - NP40
Specification
U2OS
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
Fixative
Formaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jul 02 2021

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Colon)
Antigen retrieval step
Heat mediated
Permeabilization
Yes - PBS-Tween
Specification
Colon
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
Formaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Feb 03 2021

1-10 of 84 Abreviews or Q&A

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