04-835 | Anti-dimethyl-Histone H3 (Lys79) Antibody, clone NL59, rabbit monoclonal

04-835
100 µL  
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      Overview

      Replacement Information

      Key Spec Table

      Species ReactivityKey ApplicationsHostFormatAntibody Type
      H, VrtWB, DB, Mplex, ChIP-seq, ChIPRbCulture SupernatantMonoclonal Antibody
      Description
      Catalogue Number04-835
      Replaces05-835
      Brand Family Upstate
      Trade Name
      • Upstate
      DescriptionAnti-dimethyl-Histone H3 (Lys79) Antibody, clone NL59, rabbit monoclonal
      Alternate Names
      • H3K79me2
      • Histone H3 (di methyl K79)
      Background InformationHistone 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. Dimethylation of Lys79 on histone H3 is associated with active gene transcription and is a marker for euchromatin. Histone H3 is methylated on Lys79 by the histone methyltransferase Dot1L..
      References
      Product Information
      FormatCulture Supernatant
      PresentationCulture supernantant in 0.05% sodium azide
      Applications
      ApplicationAnti-dimethyl-Histone H3 (Lys79) Antibody, clone NL59 is a rabbit monoclonal antibody for detection of dimethyl-Histone H3 (Lys79) also known as H3K79me2, Histone H3 (di methyl K79) & has been validated in ChIP, WB, DB, Mplex.
      Key Applications
      • Western Blotting
      • Dot Blot
      • Multiplexing
      • ChIP-seq
      • Chromatin Immunoprecipitation (ChIP)
      Application NotesChromatin 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
      Please refer to the EZ-Magna ChIP™ A (Cat. # 17-408) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details.

      Chromatin Immunoprecipitation: A representative lot of this antibody has been shown by an independent laboratory to work in ChIP.

      ChIP-Seq Analysis:
      A representative lot of this antibody was used by an independent laboratory for ChIP-Seq. See Egelhofer, T.A., et al. (2011). See Easwaran, H., et al. (2012).See Suzuki, H., et al. (2011).

      Dot Blotting: 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

      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 Cat. No 04-835-S, Anti-dimethyl Histone H3 (Lys79) Antibody, clone NL59 (1:500 dilution). Proteins were visualized using a Donkey anti-rabbit IgG conjugated to HRP and a chemiluminescence detection system.
      Biological Information
      ImmunogenKLH-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.
      EpitopeLys79
      CloneNL59
      HostRabbit
      SpecificityRecognizes dimethyl-histone H3 (Lys79).
      IsotypeIgG
      Species Reactivity
      • Human
      • Vertebrates
      Species Reactivity NoteBroad species cross-reactivity expected due to sequence homology
      Antibody TypeMonoclonal Antibody
      Entrez Gene Number
      Entrez Gene SummaryHistones 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.
      Gene Symbol
      • H3F3A
      • H3F3B
      Modifications
      • Methylation
      UniProt SummaryFUNCTION: 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 Weight17 kDa
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality Assuranceroutinely evaluated by immunoblot of acid extracted proteins from HeLa cells
      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 ConditionsStable for 1 year at -20°C from date of receipt.
      Handling Recommendations: Upon receipt, 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 Size100 µL
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      SDS

      Title

      Safety Data Sheet (SDS) 

      Certificates of Analysis

      TitleLot Number
      Anti-dimethyl-Histone H3 (Lys79), clone NL59 - 2142196 2142196
      Anti-dimethyl-Histone H3 (Lys79), clone NL59 - 2165866 2165866
      Anti-dimethyl-Histone H3 (Lys79), clone NL59 - 2387821 2387821
      Anti-dimethyl-Histone H3 (Lys79), clone NL59 - 2460204 2460204
      Anti-dimethyl-Histone H3 (Lys79), clone NL59 - 2049824 2049824
      Anti-dimethyl-Histone H3 (Lys79), clone NL59 - 2211076 2211076
      Anti-dimethyl-Histone H3 (Lys79), clone NL59 - 2280417 2280417
      Anti-dimethyl-Histone H3 (Lys79), clone NL59 - 2332536 2332536
      Anti-dimethyl-Histone H3 (Lys79), clone NL59 - DAM1527889 DAM1527889
      Anti-dimethyl-Histone H3 (Lys79), clone NL59 - JBC1787295 JBC1787295

      References

      Reference overviewPub Med ID
      Methylation of histone H3 on lysine 79 associates with a group of replication origins and helps limit DNA replication once per cell cycle.
      Fu, H; Maunakea, AK; Martin, MM; Huang, L; Zhang, Y; Ryan, M; Kim, R; Lin, CM; Zhao, K; Aladjem, MI
      PLoS genetics  9  e1003542  2013

      Show Abstract
      23754963 23754963
      Coordinate changes in histone modifications, mRNA levels, and metabolite profiles in clonal INS-1 832/13 β-cells accompany functional adaptations to lipotoxicity.
      Malmgren, S; Spégel, P; Danielsson, AP; Nagorny, CL; Andersson, LE; Nitert, MD; Ridderstråle, M; Mulder, H; Ling, C
      The Journal of biological chemistry  288  11973-87  2013

      Show Abstract
      23476019 23476019
      DNA methylation and histone modifications modulate the β1,3 galactosyltransferase β3Gal-T5 native promoter in cancer cells.
      Anna Caretti,Silvia M Sirchia,Silvia Tabano,Aida Zulueta,Fabio Dall'olio,Marco Trinchera
      The international journal of biochemistry & cell biology  44  2012

      Show Abstract
      22001559 22001559

      Technical Info

      Title
      White Paper - The Message in the Marks: Deciphering Cancer Epigenetics

      Posters

      Title
      Histone Modifications

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      Life Science Research > Antibodies and Assays > Primary Antibodies