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48-602MAG
Buffer Detection Kit for Magnetic Beads
1 Kit
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17-10125
Sigma-AldrichChIPAb+™ Dimethyl-Histone H3 (Lys79) - ChIP Validated Antibody and Primer Set, rabbit monoclonal
This ChIPAb+ Dimethyl-Histone H3 (Lys79) -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
More>>This ChIPAb+ Dimethyl-Histone H3 (Lys79) -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers. Less<<
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ChIPAb+™ Dimethyl-Histone H3 (Lys79) - ChIP Validated Antibody and Primer Set, rabbit monoclonal
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+ Dimethyl-Histone H3 (Lys79) set includes the Dimethyl-Histone H3 (Lys79) antibody, a negative control rabbit supernatant, and qPCR primers which amplify a 166 bp region of human GAPDH. The Dimethyl-Histone H3 (Lys79) and negative controls are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of Dimethyl-Histone H3 (Lys79)-associated chromatin.
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.
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.
References
Product Information
Format
Culture Supernatant
Control
Includes negative control rabbit supernatant and primers specific for human GAPDH.
Presentation
Anti-Dimethyl-Histone H3 (Lys79) (rabbit Monoclonal), Part No. CS204341. One vial containing 100 µL of cultured supernantant in 0.05% sodium azide. Store at -20°C. Negative Control Supernatant, Part No. CS200567. One vial containing 100 µL of cultured supernatant in 0.05% sodium azide. Store at -20°C. Control Primers, Part No. 22-004. One vial containing 75 μL of 5 μM of each primer specific for human GAPDH. Store at -20°C. FOR: TAC TAG CGG TTT TAC GGG CG REV: TCG AAC AGG AGG AGC AGA GAG CGA
This ChIPAb+ Dimethyl-Histone H3 (Lys79) -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 either 4 µL of Negative Control Supernatant , or 4 µL of Anti-dimethyl-Histone H3 (Lys79) and the Magna ChIP™ A Kit (Cat. # 17-610). Successful immunoprecipitation of dimethyl-Histone H3 (Lys79) associated DNA fragments was verified by qPCR using Control Primers as a positive locus, and human β-globin 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™ A (Cat. # 17-408) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details.
Western Blot Analysis: Representative lot data. HeLa cell nuclear extracts were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-dimethyl-Histone H3 (Lys79) (1:4,000 dilution). Proteins were visualized using a goat anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates dimethylhistone H3 (~17 kDa) (Figure 3).
Dot Blot Analysis: Absurance Histone H3 Antibody Specificity Array (Cat. No. 16-667) and Absurance Histone H2A, H2B, H4 Antibody Specificity Array (Cat. No. 16-665), which contain histone peptides with various modifications were probed with dimethyl-Histone H3 (Lys79) Antibody, clone NL59 at 1:500 dilution. Proteins were visualized using a Donkey anti-rabbit IgG conjugated to HRP and a chemiluminescence detection system.
Dot Blot: Representative lot data. Specificity of a representative lot confirmed by the ability of a 1:2500 dilution of the antibody to recognize peptides corresponding to regions of histone H3 with various modifications (Figure 4). Multiplex Assay: Representative lot data.
Biological Information
Immunogen
KLH-conjugated synthetic peptide containing the sequence DFme2KTD corresponding to dimethyl-lysine at position 79 of histone H3. The immunizing sequence is identical in human, mouse, rat and yeast. A C-terminal cysteine was added to facilitate conjugation.
Epitope
Dimethyl Lys79
Clone
NL59
Host
Rabbit
Specificity
Recognizes dimethyl-histone H3 (Lys79).
Isotype
IgG
Species Reactivity
Human
Vertebrates
Species Reactivity Note
Broad species cross-reactivity expected due to 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.
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: Sonicated chromatin prepared from HeLa cells (1 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using either 4 µL of Negative Control Supernatant , or 4 µL of Anti-dimethyl-Histone H3 (Lys79) and the Magna ChIP™ A Kit (Cat. # 17-610). Successful immunoprecipitation of dimethyl-Histone H3 (Lys79) associated DNA fragments was verified by qPCR using Control Primers (Figure 1). Please refer to the EZ-Magna ChIP™ A (Cat. # 17-408) 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. Handling Recommendations: Upon first thaw, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. 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: ~4 μL of antibody per chromatin immunoprecipitation (dependent upon biological context).
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Bestellnummer
GTIN
17-10125
04053252376535
Documentation
ChIPAb+™ Dimethyl-Histone H3 (Lys79) - ChIP Validated Antibody and Primer Set, rabbit monoclonal 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 >>
