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Mouse Monoclonal H3 di methyl K9 antibody. Suitable for Dot, ICC/IF, ChIP, ELISA, WB, IHC-P and reacts with Human, Cow, Drosophila melanogaster, Arabidopsis thaliana, Rice samples. Cited in 961 publications.


Images

Key facts

Isotype

IgG2a

Host species

Mouse

Storage buffer

pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.
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Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
DotICC/IFChIPELISAWBIHC-P
Human
Tested
Tested
Tested
Tested
Tested
Tested
Mouse
Predicted
Predicted
Predicted
Predicted
Predicted
Predicted
Rat
Predicted
Predicted
Predicted
Predicted
Predicted
Predicted
Arabidopsis thaliana
Predicted
Predicted
Expected
Predicted
Predicted
Predicted
Caenorhabditis elegans
Predicted
Predicted
Predicted
Predicted
Predicted
Predicted
Chicken
Predicted
Predicted
Predicted
Predicted
Predicted
Predicted
Common marmoset
Predicted
Predicted
Predicted
Predicted
Predicted
Predicted
Corn
Predicted
Predicted
Predicted
Predicted
Predicted
Predicted
Cow
Expected
Expected
Expected
Expected
Tested
Expected
Drosophila melanogaster
Expected
Expected
Expected
Expected
Tested
Expected
Rice
Predicted
Predicted
Expected
Predicted
Predicted
Predicted
Saccharomyces cerevisiae
Predicted
Predicted
Predicted
Predicted
Predicted
Predicted
Schizosaccharomyces pombe
Predicted
Predicted
Predicted
Predicted
Predicted
Predicted
Sheep
Predicted
Predicted
Predicted
Predicted
Predicted
Predicted
Xenopus laevis
Predicted
Predicted
Predicted
Predicted
Predicted
Predicted

Tested
Tested

Species

Human

Dilution info

1/1000

Notes

ab1220 has some cross-reactivity with H3R8me2 in peptide dot blot. Further optimization is needed to minimize the cross-reactivity in assays similar to peptide dot blot.

Expected
Expected

Species

Cow, Drosophila melanogaster

Dilution info

Use at an assay dependent concentration.

Notes

-

Predicted
Predicted

Species

Rice, Arabidopsis thaliana, Mouse, Rat, Sheep, Chicken, Saccharomyces cerevisiae, Xenopus laevis, Caenorhabditis elegans, Schizosaccharomyces pombe, Corn, Common marmoset

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

5 µg/mL

Notes

-

Expected
Expected

Species

Cow, Drosophila melanogaster

Dilution info

Use at an assay dependent concentration.

Notes

-

Predicted
Predicted

Species

Rice, Arabidopsis thaliana, Mouse, Rat, Sheep, Chicken, Saccharomyces cerevisiae, Xenopus laevis, Caenorhabditis elegans, Schizosaccharomyces pombe, Corn, Common marmoset

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

2.00000-4.00000 µg chromatin for 25.00000 µg chromatin

Notes

Use AFM ChIP primer pair ab269259 as positive control.

Expected
Expected

Species

Cow

Dilution info

-

Notes

-

Species

Drosophila melanogaster

Dilution info

-

Notes

-

Species

Arabidopsis thaliana

Dilution info

2.00000-4.00000 µg for 25.00000 µg chromatin

Notes

Use AFM ChIP primer pair ab269259 as positive control.

Species

Rice

Dilution info

2.00000-4.00000 µg for 25.00000 µg chromatin

Notes

Use AFM ChIP primer pair ab269259 as positive control.

Predicted
Predicted

Species

Mouse, Rat, Sheep, Chicken, Saccharomyces cerevisiae, Xenopus laevis, Caenorhabditis elegans, Schizosaccharomyces pombe, Corn, Common marmoset

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

0.00025-1.00000 µg/mL

Notes

when testing with immunogen peptide.

Expected
Expected

Species

Cow, Drosophila melanogaster

Dilution info

Use at an assay dependent concentration.

Notes

-

Predicted
Predicted

Species

Rice, Arabidopsis thaliana, Mouse, Rat, Sheep, Chicken, Saccharomyces cerevisiae, Xenopus laevis, Caenorhabditis elegans, Schizosaccharomyces pombe, Corn, Common marmoset

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1.00000-5.00000 µg/mL

Notes

-

Species

Cow

Dilution info

1.00000-5.00000 µg/mL

Notes

-

Species

Drosophila melanogaster

Dilution info

1.00000-5.00000 µg/mL

Notes

-

Predicted
Predicted

Species

Rice, Arabidopsis thaliana, Mouse, Rat, Sheep, Chicken, Saccharomyces cerevisiae, Xenopus laevis, Caenorhabditis elegans, Schizosaccharomyces pombe, Corn, Common marmoset

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Expected
Expected

Species

Cow, Drosophila melanogaster

Dilution info

Use at an assay dependent concentration.

Notes

-

Predicted
Predicted

Species

Rice, Arabidopsis thaliana, Mouse, Rat, Sheep, Chicken, Saccharomyces cerevisiae, Xenopus laevis, Caenorhabditis elegans, Schizosaccharomyces pombe, Corn, Common marmoset

Dilution info

-

Notes

-

Target data

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.

Alternative names

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Mouse Monoclonal H3 di methyl K9 antibody. Suitable for Dot, ICC/IF, ChIP, ELISA, WB, IHC-P and reacts with Human, Cow, Drosophila melanogaster, Arabidopsis thaliana, Rice samples. Cited in 961 publications.

Alternative names

Key facts

Isotype

IgG2a

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

mAbcam 1220

Purification technique

Affinity purification Protein G

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.

Light chain type

kappa

Concentration
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Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

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.

Abcam is leading the way to address reproducibility in scientific research with our highly validated recombinant monoclonal and recombinant multiclonal antibodies. Search & select one of Abcam's thousands of recombinant alternatives to eliminate batch-variability and unnecessary animal use.

If you do not find a host species to meet your needs, our catalogue and custom Chimeric range provides scientists the specificity of Abcam's RabMAbs in the species backbone of your choice. Remember to also review our range of edited cell lines, proteins and biochemicals relevant to your target that may help you further your research goals.

Abcam antibodies are extensively validated in a wide range of species and applications, so please check the reagent specifications meet your scientific needs before purchasing. If you have any questions or bespoke requirements, simply visit the Contact Us page to send us an inquiry or contact our Support Team ahead of purchase.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

12 product images

  • ChIP - Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220), expandable thumbnail
    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/

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

    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).

  • Western blot - Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220), expandable thumbnail
    This image is courtesy of an anonymous customer review

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

    All lanes: Western blot - 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: 15 kDa

    Observed band size: 15 kDa

    Exposure time: 1min

  • ChIP - Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220), expandable thumbnail
    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/

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

    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), expandable thumbnail
    This image is courtesy of an anonymous customer review.

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

    Blocking buffer: 5% milk

    All lanes: Western blot - Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (AB1220) at 1/1000 dilution

    All lanes: HEK293 at 30 µg

    Secondary

    All lanes: HRP conjugated polyclonal Sheep anti-Mouse IgG at 1/5000 dilution

    Performed under reducing conditions.

    Predicted band size: 15 kDa

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

    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), expandable thumbnail
    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 - Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)

    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), expandable thumbnail

    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 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
    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

    All lanes: Western blot - Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (AB1220)

    All lanes: Calf thymus histone lysate

    Secondary

    All lanes: Western blot - Rabbit Anti-Mouse IgG H&L (HRP) (AB6728) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 15 kDa

    Exposure time: 1min

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

    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), expandable thumbnail

    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), expandable thumbnail

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

    Lane 1 : Marker.

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

    All lanes: Western blot - Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (AB1220) at 1 µg/mL

    All lanes: HeLa (Human epitherlial carcinoma cell line) Whole Cell Lysate at 20 µg

    Secondary

    All lanes: 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: 15 kDa

    Observed band size: 17 kDa

    Exposure time: 5min

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

    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).

  • Dot Blot - Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220), expandable thumbnail

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

    Dot blot analysis of Histone H3 (di methyl K9) with ab1220 at 1/1000 dilution (0.889 μg/mL) followed by Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution.

    Lane 1: Human Histone H3 peptide
    Lane 2: Human Histone H3 (Mono-Methylation R8) peptide
    Lane 3: Human Histone H3 (Di-Methylation R8) peptide
    Lane 4: Human Histone H3 (Tri-Methylation R8) peptide
    Lane 5: Human Histone H3 (Mono-Methylation K9) peptide
    Lane 6: Human Histone H3 (Di-Methylation K9) peptide
    Lane 7: Human Histone H3 (Tri-Methylation K9) peptide
    Lane 8: Human Histone H3 (Mono-Methylation K4) peptide
    Lane 9: Human Histone H3 (Di-Methylation K4) peptide
    Lane 10: Human Histone H3 (Tri-Methylation K4) peptide

    ab1220 has some cross-reactivity with H3R8me2 in peptide dot blot. Further optimization is needed to minimize the cross-reactivity in assays similar to peptide dot blot

    Blocking and dilution buffer: 5% NFDM/TBST.

    Exposure time: 180 seconds

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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