Key features and details
- Rabbit polyclonal to Histone H3 (tri methyl K9) - ChIP Grade, purified
- Suitable for: Dot blot, ChIP, WB, ICC/IF, ChIP-sequencing
- Reacts with: Human
- Isotype: IgG
Product nameAnti-Histone H3 (tri methyl K9) antibody - ChIP Grade, purified
See all Histone H3 primary antibodies
DescriptionRabbit polyclonal to Histone H3 (tri methyl K9) - ChIP Grade, purified
Tested applicationsSuitable for: Dot blot, ChIP, WB, ICC/IF, ChIP-sequencingmore details
Species reactivityReacts with: Human
Synthetic peptide corresponding to Human Histone H3 (tri methyl K9) conjugated to keyhole limpet haemocyanin.
- ICC/IF: HeLa cells. ChIP: HeLa cells. WB: Histone extracts from HeLa cells.
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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 long term. Avoid freeze / thaw cycle.
Storage bufferPreservatives: 0.05% Sodium azide, 0.05% Proclin 300
Constituent: 99% PBS
Concentration information loading...
PurityProtein A purified
Our Abpromise guarantee covers the use of ab195497 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ChIP||Use at an assay dependent concentration.
|WB||1/1000. Predicted molecular weight: 15 kDa.|
|ChIP-sequencing||Use at an assay dependent concentration.
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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ChIP analysis using Human HeLa cells, labeling Histone H3 (tri methyl K9) with ab195497 and optimized PCR primer sets for qPCR. ChIP was performed using sheared chromatin from 1000,000 cells. A titration of the antibody consisting of 1, 2, 5 and 10 μg per ChIP experiment was analysed. IgG (2 μg/IP) was used as negative IP control. QPCR was performed with primers for the heterochromatin marker Sat2 and for the ZNF510 gene, used as positive controls, and for the promoters of the active c-fos and GAPDH genes, used as negative controls. The graph shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).
ChIP analysis using HeLa cells, labeling Histone H3 (tri methyl K27) with ab195497. ChIP was performed using sheared chromatin from 1000,000 cells. The IP’d DNA was analysed by QPCR with optimized PCR primer pairs for the promoter regions of the active GAPDH and EIF4A2 genes, for the coding region of the ZNF510 gene and for the Sat2 satellite repeat. The graph shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).
ChIP-seq analysis using Human HeLa cells, labeling Histone H3 (tri methyl K9) with ab195497 at 0.5 µg and optimized PCR primer sets for qPCR as described in above. The 36 bp tags were aligned to the human genome using the BWA algorithm. The peak distribution is shown along the long arm of chromosome 19 and a zoomin to an enriched region containing several ZNF repeat genes (A and B) and the enrichment at ZNF510 and ZNF12 on chromosome 9 and 7 respectively (C and D).
Dot Blot analysis of peptides containing modified and unmodified sequences of histone H3 and H4, labeling Histone H3 (tri methyl K9) with ab195497 at 1/20000 dilution. 0.2-100 pmol of the peptide containing the respective histone modification were spotted onto the membrane.
All lanes : Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade, purified (ab195497) at 1/1000 dilution (in TBS-Tween containing 5% skimmed milk)
Lane 1 : Histone extracts from HeLa cells at 15 µg
Lane 2 : Recombinant H3 at 1 µg
Predicted band size: 15 kDa
Immunofluorescent analysis of HeLa cells labeling Histone H3 (tri methyl K9) with ab195497 at 1/500 dilution in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488 (left), DAPI (center) or a merge of the two stainings (right). Cells were fixed with 4% formaldehyde for 10 minutes and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA.
ab195497 has not yet been referenced specifically in any publications.