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48-602MAG
Buffer Detection Kit for Magnetic Beads
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Anti-dimethyl (Lys4) dimethyl (Lys9) Histone H3 Antibody is a rabbit polyclonal antibody for detection of dimethyl (Lys4) dimethyl (Lys9) Histone H3 also known as H3K4me2K9me2 has been validated in ChIP, WB, Mplex.
More>>Anti-dimethyl (Lys4) dimethyl (Lys9) Histone H3 Antibody is a rabbit polyclonal antibody for detection of dimethyl (Lys4) dimethyl (Lys9) Histone H3 also known as H3K4me2K9me2 has been validated in ChIP, WB, Mplex. Less<<
SDB (Sicherheitsdatenblätter), Analysenzertifikate und Qualitätszertifikate, Dossiers, Broschüren und andere verfügbare Dokumente.
Histone H3 is one of the five main histone proteins involved in the structure of chromatin in eukaryotic cells. Featuring a main globular domain and a long N-terminal tail, H3 is involved with the structure of the nucleosomes of the 'beads on a string' structure. The N-terminal tail of histone H3 protrudes from the globular nucleosome core and can undergo several different types of epigenetic modifications that influence cellular processes. These modifications include the covalent attachment of methyl or acetyl groups to lysine and arginine amino acids and the phosphorylation of serine or threonine. Acetylation of histone H3 occurs at several different lysine positions in the histone tail and is performed by a family of enzymes known as Histone Acetyl Transferases (HATs). Acetylation of lysine14 is commonly seen in genes that are being actively transcribed into RNA.
References
Product Information
Format
Affinity Purified
Control
HeLa nuclear extract
Presentation
Affinity purified rabbit polyclonal antibody in buffer containing 0.1 M Tris-Glycine (pH7.4), 150 mM NaCl with 0.05% sodium azide.
Applications
Application
Anti-dimethyl (Lys4) dimethyl (Lys9) Histone H3 Antibody is a rabbit polyclonal antibody for detection of dimethyl (Lys4) dimethyl (Lys9) Histone H3 also known as H3K4me2K9me2 has been validated in ChIP, WB, Mplex.
Key Applications
Western Blotting
Multiplexing
Chromatin Immunoprecipitation (ChIP)
Application Notes
Chromatin Immunoprecipitation: An independent lab has shown that this antibody performs in chromatin immunoprecipitation (ChIP).
Biological Information
Immunogen
Synthetic peptide corresponding to residues surrounding and including dimethlyated Lys 4 and dimethlyated Lys9 of Histone H3.
Epitope
Dimethylated Lys4 & Lys9
Concentration
Please refer to the Certificate of Analysis for the lot-specific concentration.
Host
Rabbit
Specificity
This antibody only detects Histone H3 when dimethylated on both Lysine 4 and Lys9.
Species Reactivity
Human
Vertebrates
Species Reactivity Note
Human tested. Expected to react with most other species based on extreme conserved homology among 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 STRUCTURE: 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.
Ubiquitinated By similarity.
SEQUENCE 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
Evaluated by Western Blotting on HeLa acid extracted nuclear preps (positive) and recombinant Histone H3 (negative).
Western Blotting Analysis: 1:500 dilution of this antibody was used to dimethyl (Lys4) dimethyl (Lys9) Histone H3 in acid extracted HeLa nuclear extracts.
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.
Immune cells develop endotoxin tolerance (ET) after prolonged stimulation. ET increases the level of a repression mark H3K9me2 in the transcriptionally silent chromatin specifically associated with pro-inflammatory genes. However, it is not clear what proteins are functionally involved in this process. Here we show that a novel chromatin activity-based chemoproteomic (ChaC) approach can dissect the functional chromatin protein complexes that regulate ET-associated inflammation. Using UNC0638 that binds the enzymatically active H3K9-specific methyltransferase G9a/GLP, ChaC reveals that G9a is constitutively active at a G9a-dependent mega-dalton repressome in primary endotoxin-tolerant macrophages. G9a/GLP broadly impacts the ET-specific reprogramming of the histone code landscape, chromatin remodelling and the activities of select transcription factors. We discover that the G9a-dependent epigenetic environment promotes the transcriptional repression activity of c-Myc for gene-specific co-regulation of chronic inflammation. ChaC may also be applicable to dissect other functional protein complexes in the context of phenotypic chromatin architectures.
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 >>