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17-675
Sigma-AldrichChIPAb+ Histone H3 (Unmodified Lys4) - ChIP Validated Antibody and Primer Set
This ChIPAb+ Histone H3 (Unmodified Lys4) -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
More>>This ChIPAb+ Histone H3 (Unmodified Lys4) -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers. Less<<
MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
ChIPAb+ Histone H3 (Unmodified Lys4) - 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+ Histone H3 (Unmodified Lys4) set includes the Anti-Histone H3 (unmodified Lys4) antibody, a negative control antibody (purified Mouse IgG), and qPCR primers which amplify a 110 bp region within the promoter of the human β-globin gene. The histone H3 (Unmodified Lys4) and negative control antibodies are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of histone H3 (Unmodified Lys4)-associated chromatin.
Alternate Names
H3 histone family, member T
histone 3
H3
histone cluster 3
H3
References
Product Information
Format
Protein G Purified
Control
Includes negative control mouse IgG antibody and control primers specific for human β-globin promoter.
Presentation
Anti-Histone H3 (Unmodified Lys4) (mouse monoclonal IgG2bκ, clone CMA301). One vial containing 50 μg of protein G purified antibody in 50 μL PBS containing 0.05% sodium azide. 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 β-Globin. One vial containing 75 μL of 5 μM of each primer specific for the human β-globin promoter. Store at -20°C. FOR: AGG ACA GGT ACG GCT GTC ATC REV: TTT ATG CCC AGC CCT GGC TC
This ChIPAb+ Histone H3 (Unmodified Lys4) -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
Key Applications
Western Blotting
Chromatin Immunoprecipitation (ChIP)
Application Notes
Chromatin Immunoprecipitation: Sonicated chromatin prepared from HeLa cells (2 X 106 cell equivalents per IP) was subjected to chromatin immunoprecipitation using 2 µg of either normal mouse IgG or Anti- Histone H3 (Unmodified Lys4) antibody and the Magna ChIP G Kit (Cat. #17-611). Successful immunoprecipitation of histone H3 associated DNA fragments was verified by qPCR using β-globin ChIP Primers versus GAPDH Promoter Control Primers. Data is presented as percent input of each IP sample relative to input chromatin, with the indicated primer sets (Please see figures).
Western Blot Analysis: Acid extracted proteins from HeLa cells (Lane 1) and Recombinant Histones (Lane 2-6) were resolved by electrophoresis, transferred to PVDF membrane and probed with anti-Histone H3 (Unmodified Lys4) (0.1 μg/ml). Proteins were visualized using a goat-anti mouse secondary antibody conjugated to HRP (Cat. #AP124P) and a chemiluminescence detection system (Please see figures).
Biological Information
Immunogen
The histone H3 (Unmodified Lys4) purified antibody is made against a synthetic peptide corresponding to amino acids 1-12 of histone H3.
Epitope
a.a. 1-12
Clone
CMA301
Host
Mouse
Specificity
Recognizes unmodified histone H3 (Lys4), Mr 17 kDa. Epitope specificity of antibody as determined by ELISA maps near Lys4. Antibody tolerates modifications on Lys9 and Ser10 but not Lys4 or Thr31.
Isotype
IgG2bκ
Species Reactivity
Vertebrates
Species Reactivity Note
Human. The peptide sequence is identical in a wide range of animal and plant species, so broad cross-reactivity is expected.
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. 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. 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. SIMILARITY: Belongs to the histone H3 family. SEQUENCE CAUTION: The sequence CAH73371.1 differs from that shown. Reason: Erroneous gene model prediction.
Molecular Weight
Histone H3 at ~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-Histone H3 (Unmodified Lys4) antibody and the Magna ChIP G Kit (Cat. #17-611). Successful immunoprecipitation of histone H3 (Unmodified Lys4) associated DNA fragments was verified by qPCR using β-Globin ChIP Primers (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
Catalog Number
GTIN
17-675
04053252674808
Documentation
ChIPAb+ Histone H3 (Unmodified Lys4) - ChIP Validated Antibody and Primer Set SDS
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. Learn More >>
