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Histone H3 (K9me3) polyclonal antibody

  • Catalog # : PAB31325
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  • Specification
  • Product Description:
  • Rabbit polyclonal antibody raised against synthetic peptide of Histone H3 (K9me3).
  • Immunogen:
  • A synthetic peptide (conjugated with KLH) corresponding to Histone H3, trimethylated at lysine 9.
  • Host:
  • Rabbit
  • Reactivity:
  • Human, Mouse
  • Form:
  • Liquid
  • Purification:
  • Affinity purification
  • Recommend Usage:
  • ELISA (1:5000)
    Western Blot (1:1000)
    ChIP (1 ug/IP)
    Dot Blot (1:20000)
    Immunofluorescence (1:500)
    The optimal working dilution should be determined by the end user.
  • Storage Buffer:
  • In PBS (0.05% sodium azide, 0.05% proclin 300).
  • Storage Instruction:
  • Store at -20°C. For long term storage store at -80°C.
    Aliquot to avoid repeated freezing and thawing.
  • Note:
  • This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
  • Publication Reference
  • 1.
  • DNA methylation heterogeneity defines a disease spectrum in Ewing sarcoma.
    Sheffield NC, Pierron G, Klughammer J, Datlinger P, Schönegger A, Schuster M, Hadler J, Surdez D, Guillemot D, Lapouble E, Freneaux P, Champigneulle J, Bouvier R, Walder D, Ambros IM, Hutter C, Sorz E, Amaral AT, de Álava E, Schallmoser K, Strunk D, Rinner B, Liegl-Atzwanger B, Huppertz B, Leithner A, de Pinieux G, Terrier P, Laurence V, Michon J, Ladenstein R, Holter W, Windhager R, Dirksen U, Ambros PF, Delattre O, Kovar H, Bock C, Tomazou EM.Nat Med. 2017 Mar;23(3):386-395.
  • 2.
  • Immunometabolic Pathways in BCG-Induced Trained Immunity.
    Arts RJ, Carvalho A, La Rocca C, Palma C, Rodrigues F, Silvestre R, Kleinnijenhuis J, Lachmandas E, Gon?alves LG, Belinha A, Cunha C, Oosting M, Joosten LA, Matarese G, van Crevel R, Netea MG.Cell Rep. 2016 Dec 6;17(10):2562-2571.
  • 4.
  • β-Glucan Reverses the Epigenetic State of LPS-Induced Immunological Tolerance.
    Novakovic B, Habibi E, Wang SY, Arts RJ, Davar R, Megchelenbrink W, Kim B, Kuznetsova T, Kox M, Zwaag J, Matarese F, van Heeringen SJ, Janssen-Megens EM, Sharifi N, Wang C, Keramati F, Schoonenberg V, Flicek P, Clarke L, Pickkers P, Heath S, Gut I, Netea MG, Martens JH, Logie C, Stunnenberg HG.Cell. 2016 Nov 17;167(5):1354-1368.e14.
  • 8.
  • Epigenetic dynamics of monocyte-to-macrophage differentiation.
    Wallner S, Schröder C, Leitão E, Berulava T, Haak C, Beißer D, Rahmann S, Richter AS, Manke T, Bönisch U, Arrigoni L, Fröhler S, Klironomos F, Chen W, Rajewsky N, Müller F, Ebert P, Lengauer T, Barann M, Rosenstiel P, Gasparoni G, Nordström K, Walter J, Brors B, Zipprich G, Felder B, Klein-Hitpass L, Attenberger C, Schmitz G, Horsthemke B.Epigenetics Chromatin. 2016 Jul 29;9:33.
  • Applications
  • ChIP
  • ChIP
  • ChIP assays were performed using human HeLa cells. A titration consisting of 0.2, 0.5, 1, 2 and 5 ug of antibody per ChIP experiment was analyzed. IgG (1 ug/IP) was used as a negative IP control.
  • ChIP
  • ChIP
  • ChIP assays were performed using human K562 cells. A titration consisting of 0.2, 0.5, 1 and 2 ug of antibody per ChIP experiment was analyzed. IgG (1 ug/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter of the active genes GAPDH, c-fos and EIF4A2, used as negative controls, and for ZNF510 and the Sat2 satellite repeat, used as positive controls. The figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).
  • ChIP-Seq
  • ChIP-Seq
  • ChIP was performed on sheared chromatin from 100,000 K562 cells using antibody. The figure shows the H3K9me3 signal distribution along the long arm of human chromosome 19 and a zoomin to an enriched region containing several ZNF repeat genes and the signal distribution in a 200 kb region from chromosome 12 surrounding the ZNF12 gene.
  • Western Blot
  • Western Blot
  • Western Blot analysis of (1) 25 ug whole cell extracts of Hela cells, (2) 15 ug histone extracts of Hela cells, (3) 1 ug of recombinant histone H2A, (4) 1 ug of recombinant histone H2B, (5) 1 ug of recombinant histone H3, (6) 1 ug of recombinant histone H4.
  • Immunofluorescence
  • Immunofluorescence
  • Immunofluorescent staining of Hela cell line with antibody followed by an anti-rabbit antibody conjugated to Alexa488 (top). The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings (bottom).
  • ELISA
  • ELISA
  • ELISA is a quantitative method used to determine the titer of the antibody using a serial dilution of antibody against Histone H3 (K9me3). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:11500.
  • Dot Blot
  • Dot Blot
  • Cross reactivity test using the Histone H3 (K9me3) antibody.
    Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K9. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20000. The figure shows a high specificity of the antibody for the modification of interest.
  • Application Image
  • Gene Information
  • Entrez GeneID:
  • 8350
  • Gene Name:
  • HIST1H3A
  • Gene Alias:
  • H3/A,H3FA
  • Gene Description:
  • histone cluster 1, H3a
  • Gene Summary:
  • Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. This structure consists of approximately 146 bp of DNA wrapped around a nucleosome, an 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 found in the large histone gene cluster on chromosome 6p22-p21.3. [provided by RefSeq
  • Other Designations:
  • H3 histone family, member A,histone 1, H3a
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