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

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Overview

Product name

Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade
Histone H3 抗体 (41件) 一覧

Description

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

Specificity

By peptide ELISA ab1220 recognises 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

ICC/IF, IP, WB, Flow Cyt, ELISA, IHC-P, ChIPmore details

Cross reactivity

Reacts with

Mouse, Rat, Chicken, Cow, Human, Xenopus laevis, Arabidopsis thaliana, Caenorhabditis elegans, Fruit fly (Drosophila melanogaster), Schizosaccharomyces pombe, Corn, Rice, Other

Predicted to work with

Sheep, Saccharomyces cerevisiae

Immunogen

Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Histone H3, di methylated at K9.

Immunogen の所有権に関して

. Hybridomas were prepared and the resulting clones were positively screened by ELISA against the immunising peptide conjugated to BSA. Clones were negatively screened against both the non-modified equivalent peptide and against a peptide corresponding to dimethylated K27.

Properties

Form

Liquid

Storage instructions

Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.

Storage buffer

Preservative: 0.05% Sodium Azide
Constituents: PBS, pH 7.5

Concentration

Concentration information loading...

Purity

IgG fraction

Clonality

Monoclonal

Clone number

mAbcam 1220

Isotype

IgG2a

Light chain type

kappa

Research areas

Epigenetics and Nuclear Signaling >> ChIP'ing antibodies >> ChIP'ing antibodies
Epigenetics and Nuclear Signaling >> Histones >> H3 >> Methylated

  • Western blot - Histone H3 (di methyl K9) antibody [mAbcam 1220] (ab1220)Western blot - Histone H3 (di methyl K9) antibody [mAbcam 1220] (ab1220) image (enlarge)

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

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

Applications

Show applications key

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.

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 accessionP68431 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).

Information by UniProt

Alternative names

  • Histone H3.1 antibody
  • FLJ92264 antibody
  • H3 histone antibody
  • H3 histone antibody
  • H3 histone family, member A antibody
  • H3/a antibody
  • H3/b antibody
  • H3/c antibody
  • H3/d antibody
  • h3/f antibody
  • H3/h antibody
  • H3/i antibody
  • H3/j antibody
  • H3/k antibody
  • H3/l antibody
  • H31_HUMAN antibody
  • H3F1K antibody
  • H3F3 antibody
  • H3FA antibody
  • H3FB antibody
  • H3FC antibody
  • H3FD antibody
  • H3FF antibody
  • H3FH antibody
  • H3FI antibody
  • H3FJ antibody
  • H3FK antibody
  • H3FL 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 cluster 1, H3b antibody
  • Histone cluster 1, H3c antibody
  • Histone cluster 1, H3d antibody
  • Histone cluster 1, H3e antibody
  • Histone cluster 1, H3f antibody
  • Histone cluster 1, H3g antibody
  • Histone cluster 1, H3i antibody
  • Histone cluster 1, H3j antibody
  • Histone H 3 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

Database links

see all

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

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

Western blot - Histone H3 (di methyl K9) antibody [mAbcam 1220] (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 Histone H3 peptide - mono methyl K9 (ab1771) at 1 µg/ml
Lane 4 : Calf thymus histone lysate with Histone H3 peptide - di methyl K9 (ab1772) at 1 µg/ml
Lane 5 : Calf thymus histone lysate with Histone H3 peptide - tri methyl K9 (ab1773) at 1 µg/ml
Lane 6 : Calf thymus histone lysate with Histone H3 peptide - di methyl K4 (ab7768) at 1 µg/ml
Lane 7 : Calf thymus histone lysate with Histone H3 peptide - di methyl K27 (ab1781) at 1 µg/ml

Secondary
Rabbit polyclonal Secondary Antibody to 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 - Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)

ChIP - 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.
 

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

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

Interphase HeLa cells were fixed in paraformaldehyde and labelled with ab1220 (1/500). The staining is shown in green. The cells were counterstained with DAPI (red).

This image was submitted as part of a review by Kirk McManus

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

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

ELISA using ab1220 at varying antibody concentrations.

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

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

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

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

ab1220 staining mouse embryonic stem cells by flow cytometry (gated on all living cells). The cells were trypsinized and stained with the antibody at 1ug/1.5 x 105 cells in a permeabilization buffer. A Cy3 ® conjugated goat anti-mouse antibody was used as the secondary.

This image is courtesy of an Abreview submitted by Prof Albrecht Müller

See Abreview

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

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

ab1220 staining rat liver tissue sections by IHC-P.  Sections were formaldehyde fixed and subjected to heat mediated antigen retrieval in citrate buffer pH 6.0 prior to blocking with 5% serum for 30 minutes at 20°C.  The primary antibody was diluted 1/400 and incubated with the sample for 45 minutes at 20°C.  A HRP-conjugated goat anti-mouse antibody was used as the secondary.

This image is courtesy of an anonymous Abreview

See Abreview

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

This product has been referenced in:

  • Oyer JAet al. Aberrantly silenced promoters retain a persistent memory of the silenced state after long-term reactivation. Mutat Res 706:21-7 (2011). ChIP; Mouse.Read more (PubMed: 21035468) »
  • Henzel JVet al. An asymmetric chromosome pair undergoes synaptic adjustment and crossover redistribution during Caenorhabditis elegans meiosis: implications for sex chromosome evolution. Genetics 187:685-99 (2011).Read more (PubMed: 21212235) »

See all 55 publications for this product

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

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"