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Anti-Histone H3 (tri methyl K36) antibody - ChIP Grade (ab9050)產品信息：
- Product nameAnti-Histone H3 (tri methyl K36) antibody - ChIP GradeSee all Histone H3 primary antibodies ...
- Description
Rabbit polyclonal to Histone H3 (tri methyl K36) - ChIP Grade
- SpecificityHistone H3 tri methylated at lysine 36
- Tested applicationsICC/IF, CHIPseq, WB, ChIP, ChIP/Chip, IHC-P more details
- Species reactivity
Reacts with: Mouse, Rat, Cow, Human, Saccharomyces cerevisiae, Xenopus laevis, Arabidopsis thaliana, Caenorhabditis elegans, Fruit fly (Drosophila melanogaster), Schizosaccharomyces pombe, Zebrafish, Rice, Trypanosoma brucei
Predicted to work with: Plants
- Immunogen
Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Histone H3, tri methylated at K36.
Read Abcam's proprietary immunogen policy
(Peptide available as ab1785.)
- Positive control
- Calf Thymus Histone Preparation; Hela whole cell extract
- General notes
For detection of methylated histone H3
Properties
- FormLiquid
- Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
- Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 Concentration 100 µg at 1 mg/ml
- PurityImmunogen affinity purified
- Primary antibody notes For detection of methylated histone H3
- Clonality Polyclonal
- IsotypeIgG
- Research Areas
- Epigenetics and Nuclear Signaling
- Histones
- H3
- Methylated
- Epigenetics and Nuclear Signaling
- ChIP'ing antibodies
- ChIP'ing antibodies
Applications
Our Abpromise guarantee covers the use of ab9050 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Notes |
|---|---|
| ICC/IF | ICC/IF: Use a concentration of 1 µg/ml. |
| CHIPseq | CHIPseq: Use at an assay dependent dilution. PubMed: 19581485 |
| WB | WB: Use a concentration of 1 µg/ml. Detects a band of approximay 15 kDa (predicted molecular weight: 15 kDa).Can be blocked with Human Histone H3 (tri methyl K36) peptide (ab1785). |
| ChIP | ChIP: Use 4µg for 106 cells. |
| ChIP/Chip | ChIP/Chip: Use at an assay dependent dilution. |
| IHC-P | IHC-P: Use a concentration of 0.5 - 10 µg/ml. |
Target
- 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.
- Post-translational
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
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
see all
Alternative names
- H3 histone family, member J antibody
- FLJ92264 antibody
- H3 histone antibody
see all
Anti-Histone H3 (tri methyl K36) antibody - ChIP Grade images
ChIP - Histone H3 (tri methyl K36) antibody - ChIP Grade (ab9050)
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 ab9050 (blue), and 20µl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified on the GAPDH (active) and MYO-D (inactive) promoters and over the ý-Actin gene (active). Schematic diagram of the ý-Actin gene is shown on the top of the figure. Black boxes represent exons and thin lines represent introns. PCR products are depicted as bars under the gene.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Histone H3 (tri methyl K36) antibody - ChIP Grade (ab9050)
Staining of human tonsil using ab9050 is shown. Antigen retrieval was performed using Tris EDTA at pH9. Nuclei of lymphoid cells in the interfollicular area of a human tonsil stained strongly positive as well as the endothelial cells of blood vessels.
Immunocytochemistry/ Immunofluorescence - Histone H3 (tri methyl K36) antibody - ChIP Grade (ab9050)
ICC/IF image of ab9050 stained human HeLa cells. The cells were methanol fixed (5 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab9050, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Histone H3 (tri methyl K36) antibody - ChIP Grade (ab9050)This image is courtesy of an anonymous Abreview
ab9050 staining rat liver tissue sections by IHC-P. The section was formaldehyde fixed and subjected to heat mediated antigen retrieval in pH 6.0 citrate buffer prior to being blocked with 5% serum for 30 minutes at 45°C. The primary antibody was diluted 1/500 and incubated for 45 minutes at 20°C. A HRP conjugated goat anti-rabbit was used as the secondary
See Abreview
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Histone H3 (tri methyl K36) antibody - ChIP Grade (ab9050)This image is courtesy of an anonymous Abreview
ab9050 staining mouse kidney tissue sections by IHC-P. The section was formaldehyde fixed and subjected to heat mediated antigen retrieval in pH 6.0 citrate buffer prior to being blocked with 5% serum for 30 minutes at 20°C. The primary antibody was diluted 1/500 and incubated for 45 minutes at 20°C. A HRP conjugated goat anti-rabbit was used as the secondary
See Abreview
ChIP - Histone H3 (tri methyl K36) antibody - ChIP Grade (ab9050)This image is courtesy of an anonymous Abreview
ChIP using ab9050 at the Pho4 locus in S. pombe. Chromatin extract from S.pombe cells was incubated with 5 ug of ab9050 overnight, and then incubated with Protein A beads for 1 hour. The immunoprecipitated DNA was quantified at the Pho4 locus by RT-PCR. No antibody was added to the beads control (red bars). A schematic diagram of the pho4 gene is shown below the graph. The grey box represents the open reading frame and PCR products are depicted by horizontal arrows. Please see anonymous abreview submitted on 29 October 2008 for additional details.See Abreview
Western blot - Histone H3 (tri methyl K36) antibody - ChIP Grade (ab9050)
All lanes : Anti-Histone H3 (tri methyl K36) antibody - ChIP Grade (ab9050) at 1 µg/ml
Lane 1 : Calf Thymus Histone Preparation Nuclear Lysate
Lane 2 : Calf Thymus Histone Preparation Nuclear Lysate with Histone H3 peptide - unmodified K36 at 0.5 µg/ml
Lane 3 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (mono methyl K36) peptide (ab1783) at 0.5 µg/ml
Lane 4 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (di methyl K36) peptide (ab1784) at 0.5 µg/ml
Lane 5 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (tri methyl K36) peptide (ab1785) at 0.5 µg/ml
Lane 6 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (tri methyl K37) peptide (ab24417) at 0.5 µg/ml
Lysates/proteins at 0.5 µg per lane.
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size : 15 kDa
Observed band size : 17 kDa (why is the actual band size different from the predicted?)
Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (tri methyl K36) antibody - ChIP Grade (ab9050)This image is courtesy of an anonymous Abreview
ab9050 staining Histone H3 (tri methyl K36) in Human Saos-2 cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.25% Triton in PBS and blocked with 1% BSA for 1 hour at room temperature. Samples were incubated with primary antibody (1/1000) for 1 hour. An Alexa Fluor®488-conjugated Goat anti-rabbit IgG polyclonal (1/250) was used as the secondary antibody.
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