Key features and details
- Rabbit polyclonal to Histone H3 (mono methyl K36) - ChIP Grade
- Suitable for: ICC, WB, ChIP
- Reacts with: Human
- Isotype: IgG
Product nameAnti-Histone H3 (mono methyl K36) antibody - ChIP Grade
See all Histone H3 primary antibodies
DescriptionRabbit polyclonal to Histone H3 (mono methyl K36) - ChIP Grade
SpecificitySpecific for human Histone H3 mono methyl K36. Shows partial cross-reactivity with di-methyl K36 (please see Western Blot image). This antibody may not be suitable for experiments on yeast lysate. Although the antibody is specifically blocked using the immunising peptide, customer feedback indicates that it detects a band using S. cerevisiae K36 point mutants. We welcome further customer feedback.
Tested Applications & Species
Application Species ChIPHuman ICCHuman
Synthetic peptide within Human Histone H3 aa 1-100 (mono methyl K36) conjugated to keyhole limpet haemocyanin. The exact sequence is proprietary.
(Peptide available as
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 the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.
One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.
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Storage instructionsShipped 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 bufferpH: 7.40
Preservative: 0.02% Sodium azide
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
Concentration information loading...
PurityImmunogen affinity purified
ChIP Related Products
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab9048 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
Use at an assay dependent concentration.
Use 2 µg for 25 µg of chromatin.
Use at an assay dependent concentration.
Use 2 µg for 25 µg of chromatin.
FunctionCore 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 similaritiesBelongs to the histone H3 family.
Developmental stageExpressed during S phase, then expression strongly decreases as cell division slows down during the process of differentiation.
modificationsAcetylation 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 localizationNucleus. Chromosome.
- Information by UniProt
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Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 2µg of ab9048 (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). Primers and probes are located in the first kb of the transcribed region.
All lanes : Anti-Histone H3 (mono methyl K36) antibody - ChIP Grade (ab9048) at 1/500 dilution
Lane 1 : Histone prep
Lane 2 : Histone prep with Human Histone H3 (mono methyl K36) peptide (ab1783) at 1 µg/ml
Lane 3 : Histone prep with
Human Histone H3 (di methyl K36) peptide (ab1784) at 1 µg/ml
Lane 4 : Histone prep with
Human Histone H3 (tri methyl K36) peptide (ab1785) at 1 µg/ml
Lane 5 : Histone prep with
Human Histone H3 (unmodified ) peptide (ab2623) at 1 µg/ml
Lane 6 : Histone prep with
Human Histone H3 (mono methyl K4) peptide (ab1340) at 1 µg/ml
Lysates/proteins at 0.5 µg per lane.
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/5000 dilution
Secondary ab: Alexa Fluor 680 Goat anti-rabbit IgG
g histone prep used per lane µ
Lane 1: ab9048 (Histone H3 Mono Methyl K36) 1/500
Lane 2: ab9048 (Histone H3 Mono Methyl K36) 1/500 + ab1783 (ab9048) (Histone H3 Mono Methyl K36) peptide 1
Lane 3: ab9048 (Histone H3 Mono Methyl K36) 1/500 + ab1794 (ab9049) (Histone H3 Di Methyl K36) peptide 1
Lane 4: ab9048 (Histone H3 Mono Methyl K36) 1/500 + ab1785 (ab9050) (Histone H3 Tri Methyl K36) peptide 1
Lane 5: ab9048 (Histone H3 Mono Methyl K36) 1/500 + ab2623 (Histone H3 (23-34) – unmodified) peptide 1
Lane 6: ab9048 (Histone H3 Mono Methyl K36) 1/500 + ab1340 (ab8895) (Histone H3 Mono methyl K4) peptide 1
ab9048 specifically recognise
ab9048 staining Histone H3 (mono methyl K36) in HeLa cells. The cells were fixed with 100% methanol (5 min) at room temperature, 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 with the antibody ab9048 at 1µg/ml and ab7291 (Mouse monoclonal to alpha Tubulin - Loading Control) used at a 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed, (pseudo-colored green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594) preadsorbed, (colored red), both used at a 1/1000 dilution for 1 hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43 µM for 1hour at room temperature.
Staining of interphase nuclei of Hela cells with ab9048 (green) at a working dilution of 1/500. The DNA is stained with DAPI. ab9048 appears to be more associated with heterochromatin (DAPI intense regions) than euchromatin (DAPI less intense regions).
All lanes : Anti-Histone H3 (mono methyl K36) antibody - ChIP Grade (ab9048) at 1/1400 dilution
All lanes : Whole cell lysate prepared from Drosophila BG3 cells
Lysates/proteins at 500000 cells per lane.
All lanes : HRP donkey anti-rabbit monoclonal at 1/20000 dilution
Developed using the ECL technique.
Observed band size: 17,42 kDa why is the actual band size different from the predicted?
Exposure time: 30 seconds
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
ab9048 has been referenced in 91 publications.
- Jones MA et al. Arabidopsis JMJD5/JMJ30 Acts Independently of LUX ARRHYTHMO Within the Plant Circadian Clock to Enable Temperature Compensation. Front Plant Sci 10:57 (2019). PubMed: 30774641
- Oqani RK et al. Iws1 and Spt6 Regulate Trimethylation of Histone H3 on Lysine 36 through Akt Signaling and are Essential for Mouse Embryonic Genome Activation. Sci Rep 9:3831 (2019). PubMed: 30846735
- Huang H et al. A CRISPR/Cas9 screen identifies the histone demethylase MINA53 as a novel HIV-1 latency-promoting gene (LPG). Nucleic Acids Res 47:7333-7347 (2019). PubMed: 31165872
- Wang L et al. H3K36 trimethylation mediated by SETD2 regulates the fate of bone marrow mesenchymal stem cells. PLoS Biol 16:e2006522 (2018). PubMed: 30422989
- Liu R et al. PHD finger protein 1 (PHF1) is a novel reader for histone H4R3 symmetric dimethylation and coordinates with PRMT5-WDR77/CRL4B complex to promote tumorigenesis. Nucleic Acids Res 46:6608-6626 (2018). PubMed: 29846670