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Key Specifications Table
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| H, Vrt, Yeast (S. cerevisiae) | WB, ChIP-seq, ChIP, DB | Rb | Serum | Polyclonal Antibody |
| Description | |
|---|---|
| Catalogue Number | 07-360 |
| Brand Family | Upstate |
| Trade Name |
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| Description | Anti-acetyl-Histone H3 (Lys27) Antibody |
| Alternate Names |
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| Background Information | Histone H3 (UniProt: P61830) is encoded by the HHT1 (also known as YBR010W, YBR0201, HHT2, SIN2, YNL031C, N2749) gene (Gene ID: 852295) in yeast. Histone H3 is a core component of nucleosome. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histone H3 can undergo various post-translational modifications that either activate or repression gene expression. Histone H3 can undergo mon-, di-, or trimethylation. Trimethylated H3K27 is shown to be tightly associated with inactive gene promoters and monomethylated H3K27 is associated with active promoters. The dimethylated H3K27 has a distribution pattern similar to H3K27. Histone H3 can also undergo acetylation at Lys9, 14, 18, 23, 27 and 56. Acetylation of histone H3 leads to transcriptional activation. Acetylation of histone defines the openness of chromatin as acetylated histones cannot pack as well together as deacetylated histones. H3K27ac as an important enhancer mark that can distinguish between active and poised enhancer elements and enhancer proximal genes that lack H3K27ac enrichment display lower expression levels compared with the average enhancer proximal gene. It is shown that amounts of H3K27ac is gradually declines fom the earliest pronuclear stage to 8-cell stage, corresponding to the major embryonic genome activation (EGA), followed by re-acetylation of H3K27 from the morula stage onwards accompanying the first cell lineage specification in IVF embryos. In Drosophila, H3K27ac is present at high levels in early embryos and declines after 4 hours as the level of trimethylated H3K27 increases. (Ref.: Creyghton, MP et al. ( ). Proc. Natl. Acad. Sci. USA 107(50); 21931-21936; Tie, F et al (2009). Dvelopment 136 (18); 3131-3141).. |
| Product Information | |
|---|---|
| Format | Serum |
| Control |
|
| Presentation | Rabbit polyclonal antiserum with 0.05% sodium azide and 30% glycerol. |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | Anti-acetyl-Histone H3 (Lys27) Antibody is a rabbit polyclonal antibody for detection of Histone H3 acetylated on lysine 27. Also known as Anti-H3K27ac this antibody is published in peer reviewed journals and is specificity verified by dot blot (DB) and validated in ChIP, ChIP-seq, WB, DB, Mplex. |
| Key Applications |
|
| Application Notes | ChIP-seq Analysis: Chromatin immunoprecipitation was performed using the Magna ChIP™ HiSens kit (cat# 17-10460), 2 µl of anti-acetyl-Histone H3 (Lys27) antibody (cat# 07-360), 20 µL Protein A/G beads, and 1e6 crosslinked HeLa cell chromatin followed by DNA purification using magnetic beads. Libraries were prepared from Input and ChIP DNA samples using standard protocols with Illumina barcoded adapters, and analyzed on Illumina HiSeq instrument. An excess of fourteen million reads from FastQ files were mapped using Bowtie (http://bowtie-bio.sourceforge.net/manual.shtml) following TagDust (http://genome.gsc.riken.jp/osc/english/dataresource/) tag removal. Peaks were identified using MACS (http://luelab.dfci.harvard.edu/MACS/), with peaks and reads visualized as a custom track in UCSC Genome Browser (http://genome.ucsc.edu) from BigWig and BED files. The highest 25% of peaks identified in the 07-360 dataset showed 96% overlap with peaks identified in the ENCODE H3K27Ac BROAD Histone track for HeLa S3. |
| Biological Information | |
|---|---|
| Immunogen | KLH-conjugated linear peptide corresponding to 11 amino acids from the N-terminal region of human histone H3 acetylated on Lysine 27. |
| Epitope | Lys27 |
| Concentration | Please refer to the Certificate of Analysis for the lot-specific concentration. |
| Host | Rabbit |
| Specificity | Recognizes Histone H3 acetylated on lysine 27. |
| Isotype | IgG |
| Species Reactivity |
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| Species Reactivity Note | Human. Predicted to react with Yeast based on 100% sequence homology. |
| Antibody Type | Polyclonal Antibody |
| Entrez Gene Number |
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| Entrez Gene Summary | Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Nucleosomes consist of approximately 146 bp of DNA wrapped around a histone octamer composed of pairs of each of the four core histones (H2A, H2B, H3, and H4). The chromatin fiber is further compacted through the interaction of a linker histone, H1, with the DNA between the nucleosomes to form higher order chromatin structures. This gene is intronless and encodes a member of the histone H3 family. Transcripts from this gene lack polyA tails; instead, they contain a palindromic termination element. This gene is located separately from the other H3 genes that are in the histone gene cluster on chromosome 6p22-p21.3. |
| Gene Symbol |
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| Modifications |
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| Purification Method | Unpurified |
| UniProt Number |
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| UniProt Summary | FUNCTION: SwissProt: Q16695 # 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. SIZE: 136 amino acids; 15508 Da SUBUNIT: The nucleosome is a histone octamer containing two molecules each of H2A, H2B, H3 and H4 assembled in one H3-H4 heterotetramer and two H2A-H2B heterodimers. The octamer wraps approximately 147 bp of DNA. SUBCELLULAR LOCATION: Nucleus. TISSUE SPECIFICITY: Expressed in testicular cells. DEVELOPMENTAL STAGE: Expressed during S phase, then expression strongly decreases as cell division slows down during the process of differentiation. PTM: Acetylation is generally linked to gene activation. Acetylation on Lys-10 impairs methylation at Arg-9. Acetylation on Lys-19 and Lys-24 favors methylation at Arg-18 (By similarity). & Citrullination at Arg-9 and/or Arg-18 by PADI4 impairs methylation and represses transcription (By similarity). & Asymmetric dimethylation at Arg-18 by CARM1 is linked to gene activation. Symmetric dimethylation at Arg-9 by PRMT5 is linked to gene repression (By similarity). & Methylation at Lys-5, Lys-37 and Lys-80 are linked to gene activation. Methylation at Lys-5 facilitates subsequent acetylation of H3 and H4. Methylation at Lys-80 is associated with DNA double-strand break (DSB) responses and is a specific target for TP53BP1. Methylation at Lys-10 and Lys-28 are linked to gene repression. Methylation at Lys-10 is a specific target for HP1 proteins (CBX1, CBX3 and CBX5) and prevents subsequent phosphorylation at Ser-11 and acetylation of H3 and H4. Methylation at Lys-5 and Lys-80 require preliminary monoubiquitination of H2B at Lys-120. Methylation at Lys-10 and Lys-28 are enriched in inactive X chromosome chromatin (By similarity). & Phosphorylated at Thr-4 by GSG2/haspin during prophase and dephosphorylated during anaphase. At centromeres, specifically phosphorylated at Thr-12 from prophase to early anaphase. Phosphorylated at Ser-11 during the whole mitosis. Phosphorylation at Ser-11, which is linked to gene activation, prevents methylation at Lys-10 but facilitates acetylation of H3 and H4. Phosphorylated at Ser-29 by MLTK isoform 1, RPS6KA5 or AURKB during mitosis or upon ultraviolet B irradiation (By similarity). & Phosphorylation at Ser-11 is crucial for chromosome condensation and cell-cycle progression during mitosis and meiosis. In addition phosphorylation at Ser-11 is important during interphase because it enables the transcription of genes following external stimulation, like stress or growth factors. Phosphorylation at Ser-11 is also an essential regulatory mechanism for neoplastic cell transformation. Phosphorylation at Ser-11 by AURKB/Aurora-B mediates the dissociation of HP1 proteins (CBX1, CBX3 and CBX5) from heterochromatin. & Ubiquitinated (By similarity). SIMILARITY: SwissProt: Q16695 ## Belongs to the histone H3 family. |
| Molecular Weight | 17 kDa |
| Product Usage Statements | |
|---|---|
| Quality Assurance | Evaluated by Western Blotting in acid extract of HeLa cells treated with 5 mM sodium butyrate. Western Blotting Analysis: A 1:5,000 dilution of this antibody detected acetyl-Histone H3 (Lys27) in acid extract of HeLa cells treated with 5 mM sodium butyrate.cells |
| Usage Statement |
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| Storage and Shipping Information | |
|---|---|
| Storage Conditions | Maintain for 2 years at -20°C from date of shipment. Aliquot to avoid repeated freezing and thawing. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. |
| Packaging Information | |
|---|---|
| Material Size | 100 µL |