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48-602MAG
Buffer Detection Kit for Magnetic Beads
1 Kit
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17-10242
Sigma-AldrichChIPAb+™ Trimethyl-Histone H3 (Lys9) - ChIP Validated Antibody and Primer Set
This ChIPAb+ Trimethyl-Histone H3 (Lys9) -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
More>>This ChIPAb+ Trimethyl-Histone H3 (Lys9) -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers. Less<<
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ChIPAb+™ Trimethyl-Histone H3 (Lys9) - 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+ Trimethyl-Histone H3 (Lys9) set includes the Trimethyl-Histone H3 (Lys9) antibody, a Normal mouse IgG, and control primers which amplify a 117 bp region of ChIP Primers, ZNF554. The Trimethyl-Histone H3 (Lys9) and negative controls are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of Trimethyl-Histone H3 (Lys9) -associated chromatin.
Alternate Names
H3K9me3
Histone H3 (tri methyl K9)
H3 histone family, member T
histone 3, H3
histone cluster 3, H3
Background Information
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. Histone proteins are highly post-translationally modified however Histone H3 is the most extensively modified of the five histones. Histone H3 sequence variants and variable modification states are thought to play a role in the dynamic and long term regulation of genes. Trimethylation of histone H3 on Lys9 (H3K9me3) is one of the most highly studied epigenetic marks. H3K9me3 functions in the repression of euchromatic genes, and in epigenetic control of heterochromatin assembly, most likely by acting as a recognition motif for the binding of chromatin-associated proteins, such as Swi6 or HP1α. The enzymes responsible for H3K9 trimethylation are SUV39H1 and SUV39H2.
References
Product Information
Format
Purified
Control
Includes normal mouse IgG and primers specific for human ZNF554.
Presentation
Anti-Trimethyl-Histone H3 (Lys9) (mouse monoclonal). One vial containing 25 µL of purified mouse monoclonal IgG1 in PBS with 0.05% sodium azide before the addition of glycerol to 30%. Store at -20°C. Normal Mouse IgG. One vial containing 25 µg of purified IgG in 25 µL of storage buffer containing 0.1% sodium azide. Store at -20°C. Control Primers, ZNF554. One vial containing 75 μL of 5 μM of each primer specific for the 3’ end of human ZNF554. Store at -20°C. FOR: CGG GGA AAA GCC CTA TAA AT REV: TCC ACA TTC ACT GCA TTC GT
This ChIPAb+ Trimethyl-Histone H3 (Lys9) -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: Representative lot data. Sonicated chromatin prepared from HeLa cells (1 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 1 µL of either Normal Mouse IgG,or 1 µL of Anti-Trimethyl Histone H3 (Lys9), and the Magna ChIP™ G Kit (Cat. # 17-611). Successful immunoprecipitation of trimethyl Histone H3 (Lys9) associated DNA fragments was verified by qPCR using ChIP primers specific for the human ZNF554 region as a positive locus and human GAPDH primers as a negative locus. (Figure 2). 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 ChIP™ G (Cat. # 17-409) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details. Western Blot Analysis: Representative lot data. Recombinant Histone H3 (Cat. #14-494) (Lane 1) and HeLa acid extract (Lane 2) were resolved by electrophoresis, transferred to PVDF membrane and probed with Anti-trimethyl Histone H3 (Lys9), clone CMA308 (2 μg/mL). Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates trimethyl Histone H3 (Lys9) (~17 kDa). (Figure 3).
Biological Information
Immunogen
KLH-conjugated synthetic peptide corresponding to human Histone H3 trimethylated on Lys9.
Epitope
Trimethyl Lys9
Clone
CMA308
Host
Mouse
Specificity
This antibody recognizes Histone H3 trimethylated at Lys9. Some cross-reactivity with dimethyl Lys9 is detected at higher concentrations, but the difference in specificity is >27-fold1. Phosphorylation of Ser10 interferes with antibody binding to trimethyl Lys91.
Isotype
IgG1κ
Species Reactivity
Human
Vertebrates
Species Reactivity Note
Human. Broad species cross-reactivity is expected, based on sequence homology.
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. [provided by RefSeq]
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
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Quality Assurance
Chromatin Immunoprecipitation: Representative lot data. Sonicated chromatin prepared from HeLa cells (1 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 1 µL of either Normal Mouse IgG, or 1 µL of Anti-Trimethyl Histone H3(Lys9), and the Magna ChIP™ G Kit (Cat. # 17-611). Successful immunoprecipitation of trimethyl Histone H3 (Lys9) associated DNA fragments was verified by qPCR using ChIP primers specific for the human ZNF554 region as a positive locus(Figure 1). Please refer to the EZ-Magna ChIP™ G (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 2-8°C from date of receipt. For maximum recovery of product, centrifuge the vial prior to removing the cap. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.
Packaging Information
Material Size
25 assays
Material Package
25 assays per set. Recommended use: ~1 μL of antibody per chromatin immunoprecipitation (dependent upon biological context).
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Numero di catalogo
GTIN
17-10242
04053252558252
Documentation
ChIPAb+™ Trimethyl-Histone H3 (Lys9) - ChIP Validated Antibody and Primer Set MSDS
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. Per saperne di più >>
