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17-683
Sigma-AldrichChIPAb+ Acetyl-Histone H3 (Lys27) - ChIP Validated Antibody and Primer Set
Acetyl-Histone H3 (Lys27) ChIP validated antibody & primer set including the ChIP-grade antibody & the specific control PCR primers used for chromatin immunoprecipitation of H3K27Ac.
More>>Acetyl-Histone H3 (Lys27) ChIP validated antibody & primer set including the ChIP-grade antibody & the specific control PCR primers used for chromatin immunoprecipitation of H3K27Ac. Less<<
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ChIPAb+ Acetyl-Histone H3 (Lys27) - ChIP Validated Antibody and Primer Set
Overview
All ChIPAb+ antibodies are individually validated for chromatin precipitation, every lot, every time. Each ChIPAb+ antibody set includes control primers (tested every lot by qPCR) to biologically validate your IP results in a locus-specific context. The qPCR protocol and primer sequences are provided, allowing researchers to validate ChIP protocols when using our antibody in their chromatin context. Each set also includes a negative control antibody to ensure specificity of the ChIP reaction. The ChIPAb+ Acetyl-Histone H3 (Lys27) set includes the Anti-acetyl-Histone H3 (Lys27) antibody, a negative control antibody (purified Mouse IgG), and qPCR primers which amplify a 178 bp region within the promoter of the human RPL10 gene. The acetyl-histone H3 (Lys27) and negative control antibodies are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of acetyl-histone H3 (Lys27)-associated chromatin.
Alternate Names
H3K27Ac
Histone H3 (acetyl K27)
Histone H3K27Ac
Background Information
Lysine acetylation is a dynamic, reversible and tightly regulated protein and histone modification that plays a major role in chromatin remodeling and in the regulation of gene expression in various cellular functions. Histone acetylation is often associated with transcriptional activation and acetylation of H3K27 is found at active enhancers.
References
Product Information
Format
Purified
Control
Included negative control mouse IgG antibody and control primers specific for human RPL10 promoter.
Presentation
Anti-acetyl-Histone H3 (Lys27) (mouse monoclonal IgG1, Clone CMA309). One vial containing 50 μg of protein G purified antibody in 50 μL PBS containing 0.05% sodium. Store at -20°C.
Normal Mouse IgG. Two vials containing 25 μg purified Mouse IgG in 25 μL storage buffer containing 0.1% sodium azide. Store at -20°C.
ChIP Primers, RPL10 Promoter. One vial containing 75 μL of 5 μM of each primer specific for the promoter region of human RPL10. Store at -20°C. FOR: ACC CGT CTT CGA CAG GAC T REV: GGA ACG GAA GAC GAG AAC AG
Acetyl-Histone H3 (Lys27) ChIP validated antibody & primer set including the ChIP-grade antibody & the specific control PCR primers used for chromatin immunoprecipitation of H3K27Ac.
Key Applications
Western Blotting
Chromatin Immunoprecipitation (ChIP)
Application Notes
Chromatin Immunoprecipitation: Sonicated chromatin prepared from HeLa cells (1 X 106 cell equivalents per IP) was subjected to chromatin immunoprecipitation using 2 µg of either a normal mouse IgG or Anti-acetyl-Histone H3 (Lys27) antibody and the Magna ChIP G (Cat. #17-611) Kit. Successful immunoprecipitation of acetyl-histone H3 (Lys27)-associated DNA fragments was verified by qPCR using β-globin Promoter ChIP Primers versus RPL10 Promoter Primers (Please see figures). Data is presented as percent input of each IP sample relative to input chromatin for each amplicon and ChIP sample as indicated. Please refer to the EZ-Magna G ChIP™ (Cat. #17-408) or EZ-ChIP™ (Cat. #17-371) protocol for experimental details.
Western Blot Analysis: Acid extracts from untreated (Lane 1) and sodium-butyrate treated (Lane 2) HeLa cells were resolved by electrophoresis, transferred to PVDF membrane and probed with Anti-acetyl Histone H3 (Lys27), clone CMA309 (0.1 μg/mL). Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and a chemiluminescence detection system (Please see figures).
Biological Information
Immunogen
The acetyl-histone H3 (Lys27) purified antibody is made against a synthetic peptide (acetylated at Lys27) corresponding to amino acids 19-37 of histone H3.
Epitope
a.a. 19-37
Clone
CMA309
Host
Mouse
Specificity
Recognizes histone H3, Mr 17 kDa, acetylated at Lys27.
Isotype
IgG
Species Reactivity
Vertebrates
Species Reactivity Note
Human, although this peptide sequence is identical in a wide range of animal and plant species.
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.
FUNCTION:Variant histone H3 which replaces conventional H3 in a wide range of nucleosomes in active genes. Constitutes the predominant form of histone H3 in non-dividing cells and is incorporated into chromatin independently of DNA synthesis. Deposited at sites of nucleosomal displacement throughout transcribed genes, suggesting that it represents an epigenetic imprint of transcriptionally active chromatin. 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. Ref.14 Ref.18 Ref.22
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. Interacts with HIRA, a chaperone required for its incorporation into nucleosomes. Ref.14
SUBCELLULAR LOCATION: Nucleus.
Developmental stage Expressed throughout the cell cycle independently of DNA synthesis.
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.
Citrullination at Arg-9 and/or Arg-18 by PADI4 impairs methylation and represses transcription.
Asymmetric dimethylation at Arg-18 by CARM1 is linked to gene activation. Symmetric dimethylation at Arg-9 by PRMT5 is linked to gene repression.
Specifically enriched in modifications associated with active chromatin such as methylation at Lys-5, Lys-37 and Lys-80. 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, which are linked to gene repression, are underrepresented. 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.
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, probably DAPK3. Phosphorylation at 'Ser-11' by AURKB is crucial for chromosome condensation and cell-cycle progression during mitosis and meiosis. In addition phosphorylation at 'Ser-11' 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, which is linked to gene activation, prevents methylation at Lys-10 but facilitates acetylation of H3 and H4. Phosphorylation at Ser-11 by AURKB mediates the dissociation of HP1 proteins (CBX1, CBX3 and CBX5) from heterochromatin. Phosphorylation at 'Ser-11' is also an essential regulatory mechanism for neoplastic cell transformation. Phosphorylated at Ser-29 by MLTK isoform 1, RPS6KA5 or AURKB during mitosis or upon ultraviolet B irradiation. Phosphorylation on Ser-32 is specific to regions bordering centromeres in metaphase chromosomes. Ref.9 Ref.10 Ref.12 Ref.13 Ref.19 Ref.20 Ref.21 Ref.29
Ubiquitinated By similarity.
SIMILARITY: Belongs to the histone H3 family.
SEQUENCE CAUTION: The sequence CAH73371.1 differs from that shown. Reason: Erroneous gene model prediction.
Molecular Weight
~17 kDa
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Quality Assurance
Chromatin Immunoprecipitation: Sonicated chromatin prepared from HeLa cells (1 X 106 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µg of either a normal mouse IgG or Anti-acetyl-Histone H3 (Lys27) antibody and the Magna ChIP G (Cat. #17-611) Kit. Successful immunoprecipitation of acetyl-histone H3 (Lys27) associated DNA fragments was verified by qPCR using ChIP Primers RPL10 Promoter (Please see figures). Please refer to the EZ-Magna G ChIP™ (Cat. #17-409) or EZ-ChIP™ (Cat. #17-371) protocol for experimental details.
Usage Statement
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Storage and Shipping Information
Storage Conditions
Stable for 1 year at -20°C from date of receipt. Aliquot upon thawing, avoid freeze thaw cycles.
Packaging Information
Material Size
25 assays
Material Package
25 assays per kit, ~2μg per chromatin immunoprecipitation
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Bestellnummer
GTIN
17-683
04053252368974
Documentation
ChIPAb+ Acetyl-Histone H3 (Lys27) - ChIP Validated Antibody and Primer Set SDB
Millipore’s Histone H3 antibodies demonstrate specificity against histone H3. See below for acetyl-, methyl-, phospho- histone H3 Antibodies and Proteins, based on the expertise of Upstate & Chemicon. Weitere Informationen >>
