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Products:Epigenetics and Nuclear Signaling >> Histones >> H3 >> Methylated
Overview
Product name
Anti-Histone H3 (mono methyl K36) antibody - ChIP Grade
See all Histone H3 products (41) ...
Description
Rabbit polyclonal to Histone H3 (mono methyl K36) - ChIP Grade
Specificity
This antibody is specific for human histone H3 mono-methylated at K36. It does not react with di or tri methyl K36. However, 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
IHC-P, ICC, ICC/IF, WB, ChIP, IFmore details
Cross reactivity
Reacts with
Mouse, Cow, Human, Saccharomyces cerevisiae, Arabidopsis thaliana, Fruit fly (Drosophila melanogaster)
Predicted to work with
all Mammals
Immunogen
Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Histone H3, mono methylated at K36.
(Peptide available as ab1783.)
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
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Note: 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
Concentration information loading...
Purity
Immunogen affinity purified
Clonality
Polyclonal
Isotype
IgG
Research areas
Epigenetics and Nuclear Signaling >> ChIP'ing antibodies >> ChIP'ing antibodies
Epigenetics and Nuclear Signaling >> Histones >> H3 >> Methylated
Applications
Show applications key
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.
IHC-P: Use at an assay dependent concentration.
ICC: Use at an assay dependent dilution.
ICC/IF: Use at an assay dependent dilution. (PubMed: 19727073)
WB: 1/1000Can be blocked with Histone H3 peptide - mono methyl K36 (ab1783).
ChIP: Use a concentration of 1-4 µg/ml
IF
IF: 1/500IF: 1/500
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).
Alternative names
- Histone H3.1 antibody
- FLJ92264 antibody
- H3 histone antibody
see all
Database links
- Entrez Gene: 326601 Cow
- 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
see all
Anti-Histone H3 (mono methyl K36) antibody - ChIP Grade images:
Western blot - Histone H3 (mono methyl K36) antibody - ChIP Grade (ab9048)
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 Histone H3 peptide - mono methyl K36 (ab1783) at 1 µg/ml
Lane 3 : Histone prep with Histone H3 peptide - di methyl K36 (ab1784) at 1 µg/ml
Lane 4 : Histone prep with Histone H3 peptide - tri methyl K36 (ab1785) at 1 µg/ml
Lane 5 : Histone prep with Histone H3 peptide - unmodified (ab2623) at 1 µg/ml
Lane 6 : Histone prep with Histone H3 peptide - mono methyl K4 (ab1340) at 1 µg/ml
Lysates/proteins at 0.5 µg per lane.
Secondary
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP) (ab6721) at 1/5000 dilution
ab9048 specifically recognises Histone H3 mono methyl K36. The activity of the antibody is quenched by the addition of the blocking peptide, ab1783 (lane 2).
Immunocytochemistry - Histone H3 (mono methyl K36) antibody - ChIP Grade (ab9048)
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).
This image was submitted as part of a review by Kirk McManus, University of Columbia
ChIP - Histone H3 (mono methyl K36) antibody - ChIP Grade (ab9048)
Chromatin was prepared from U2OS cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 min. The ChIP was performed with 25 µg of chromatin, 2 µg of ab9048 (blue), and 20 µl 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Histone H3 (mono methyl K36) antibody - ChIP Grade (ab9048)
ab9048 staining Histone H3 (mono methyl K36) in Mouse pancreatic tumor tissue by Immunohistochemistry (Formalin/ PFA-fixed paraffin-embedded tissue sections). The sections were PFA-fixed and subjected to Heat mediated antigen retrieval in citrate and permeabilized with PBST prior to blocking with 5% serum for 1 hour at room temperature. The primary antibody was diluted 1/500 in PBS and incubated with the sample for 8 hours at 4°C. A Biotin-conjugated Goat anti-Rabbit polyclonal was used as the secondary antibody, diluted 1/1000.
This image is courtesy of an anonymous Abreview.
Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (mono methyl K36) antibody - ChIP Grade (ab9048)
ICC/IF image of ab9048 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab9048, 0.1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% formaldehyde fixed (10 min) Hek293, HepG2 and MCF7 cells at 0.1µg/ml, and in 100% methanol fixed (5 min) HeLa, Hek293, HepG2 and MCF7 cells at 0.1µg/ml.
Western blot - Histone H3 (mono methyl K36) antibody - ChIP Grade (ab9048)
All lanes : Anti-Histone H3 (mono methyl K36) antibody - ChIP Grade (ab9048) at 1/1400 dilution
Lane 1 : Whole cell lysate prepared from Drosophila BG3 cells
Lane 2 : Whole cell lysate prepared from Drosophila BG3 cells
Lysates/proteins at 500000 cells per lane.
Secondary
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
Image courtesy of an anonymous Abreview.
References for Anti-Histone H3 (mono methyl K36) antibody - ChIP Grade (ab9048)
This product has been referenced in:
- Tan MKet al. SCF(FBXO22) regulates histone H3 lysine 9 and 36 methylation levels by targeting histone demethylase KDM4A for ubiquitin-mediated proteasomal degradation. Mol Cell Biol 31:3687-99 (2011).Read more (PubMed: 21768309) »
- Wen Het al. Recognition of Histone H3K4 Trimethylation by the Plant Homeodomain of PHF2 Modulates Histone Demethylation. J Biol Chem 285:9322-6 (2010). ICC/IF; Human.Read more (PubMed: 20129925) »
See all 23 publications for this product
Publishing research using ab9048? 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"