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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 (64)Q&A (12)References (683)

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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)
  • 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)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - 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: WB, ELISA, IHC-P, ChIP
  • Reacts with: Cow, Human, Arabidopsis thaliana, Rice, Recombinant fragment
  • Isotype: IgG2a

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Recombinant human Histone H3 protein (Active) (ab198757)
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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 & Species

    Application Species
    ChIP
    Human
    ELISA
    Recombinant fragment
    IHC-P
    Human
    WB
    Cow
    Human
    See all applications and species data
  • Immunogen

    Synthetic peptide corresponding to Human Histone H3 aa 1-100 (di methyl K9) conjugated to keyhole limpet haemocyanin (Cysteine residue). Clones were positively screened by ELISA against the immunising peptide. Clones were negatively screened against both the non-modified equivalent peptide and against a dimethylated K27 peptide.
    (Peptide available as ab1772)

  • Positive control

    • IHC-P: Human kidney tissue; Rat liver tissue; Mouse liver P5 tissue. Flow Cyt: Mouse embryonic stem cells. WB: HeLa whole cell lysate; Calf thymus histone lysate; HEK-293 lysate; Fruit fly embryo tissue lysate. ChIP: AFM ChIP primer pair ab269259; Chromatin prepared from U2OS 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.

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

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.

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
IHC-P
Human
WB
Cow
Human
All applications
Mouse
Rat
Sheep
Chicken
Saccharomyces cerevisiae
Xenopus laevis
Caenorhabditis elegans
Drosophila melanogaster
Schizosaccharomyces pombe
Corn
Common marmoset
Other species
Application Abreviews Notes
WB (30)
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 1 - 0.00025 µg/ml. when testing with immunogen peptide.
IHC-P (6)
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
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 1 - 0.00025 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: 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).

  • 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.

  • 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)This image is a courtesy of Anonymous Abreview

    ab1220 staining Histone H3 (di methyl K9) in Mouse Tissue sections (Liver, P5) by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and blocked with 5% BSA for 1 hour at 25°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/100 in PBS/Triton 0,05%) for 3 hours at 25°C. An Alexa Fluor® 488 conjugated Goat anti-mouse (1/250) was used as the secondary antibody.

    See Abreview

  • 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

    • Datasheet
    • SDS
  • References (683)

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

    ab1220 has been referenced in 683 publications.

    • Nicolau M  et al. The plant mobile domain proteins MAIN and MAIL1 interact with the phosphatase PP7L to regulate gene expression and silence transposable elements in Arabidopsis thaliana. PLoS Genet 16:e1008324 (2020). PubMed: 32287271
    • Jang JE  et al. PERK/NRF2 and autophagy form a resistance mechanism against G9a inhibition in leukemia stem cells. J Exp Clin Cancer Res 39:66 (2020). PubMed: 32293500
    • Chen TQ  et al. EHMT2/G9a Inhibits Aortic Smooth Muscle Cell Death by Suppressing Autophagy Activation. Int J Biol Sci 16:1252-1263 (2020). PubMed: 32174799
    • Shamsi F  et al. FGF6 and FGF9 regulate UCP1 expression independent of brown adipogenesis. Nat Commun 11:1421 (2020). PubMed: 32184391
    • Jozi M  et al. Induced DNA hypomethylation by Folic Acid Deprivation in Bovine Fibroblast Donor Cells Improves Reprogramming of Somatic Cell Nuclear Transfer Embryos. Sci Rep 10:5076 (2020). PubMed: 32193457
    View all Publications for this product

    Customer reviews and Q&As

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    1-10 of 76 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

    Abcam user community

    Verified customer

    Submitted Apr 29 2013

    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

    Abcam user community

    Verified customer

    Submitted Feb 03 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 macrophage cell line)
    Gel Running Conditions
    Reduced Denaturing (15)
    Loading amount
    5 µg
    Specification
    Mouse macrophage cell line
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    Submitted Nov 18 2020

    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 (Gastric Cancer Cell Line)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    5 µg
    Specification
    Gastric Cancer Cell Line
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    Submitted Nov 18 2020

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

    Excellent
    Abreviews
    Abreviews
    Application
    ChIP
    Sample
    Mouse Cell lysate - whole cell (Mouse macrophage cell line)
    Specification
    Mouse macrophage 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: 1% Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Nov 18 2020

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

    Excellent
    Abreviews
    Abreviews
    Application
    ChIP
    Sample
    Human Cell lysate - whole cell (Cancer Cell Line)
    Negative control
    Beads only and IgG used as negative control.
    Specification
    Cancer 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: 1% Formaldehyde
    Positive control
    Cancer cell lines treated with drug for 24 hours
    Read More

    Abcam user community

    Verified customer

    Submitted Nov 17 2020

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

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (Heart)
    Gel Running Conditions
    Reduced Denaturing (4-12%)
    Loading amount
    30 µg
    Specification
    Heart
    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

    Submitted Aug 13 2020

    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
    Mouse Tissue sections (Heart)
    Permeabilization
    Yes - Tween-20
    Specification
    Heart
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Aug 11 2020

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

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Tammar wallaby Cell (ovary)
    Permeabilization
    Yes - 0.1% (v/v) Triton X-100 in PBS
    Specification
    ovary
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 27°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Mar 12 2019

    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 - nuclear (B Cells)
    Gel Running Conditions
    Reduced Denaturing (4-20%)
    Loading amount
    20 µg
    Specification
    B Cells
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    Submitted Sep 05 2018

    1-10 of 76 Abreviews or Q&A

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