Histone H3 Antibody (Tri-Methyl-Lys9) (OADC00278)

• Product Number: OADC00278
• Name: Histone H3 Antibody (Tri-Methyl-Lys9) (OADC00278)
• Conjugation: Unconjugated
• NCBI Gene Id: 8290; 8350; 8351; 8352; 8353; 8354; 8355; 8356; 8357; 8358; 8968
• Host: Fab format with human framework
• Clonality: Monoclonal
• Gene Full Name: histone cluster 3, H3
• Alias Symbols: H3t, H3.4, H3/g, H3FT, HIST3H3
• Product Format: Liquid
• Description of Target: 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 replication-dependent histone that is 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.
• Reconstitution and Storage: Store at -20C. Avoid repeated freeze/thaw cycles.
• Specificity: Selected using tailored phage-display libraries, and produced in E.coli eliminating animals from the antibody-production process. This recombinant antibody shows superior specificity and affinity as well as no lot-to-lot variation.
• Application Info: CHIP: 0.2-1.8 ug/CHIP
  IF: (Hattori T. et al., 2013)
• Uniprot ID: Q16695
• Protein Name: histone H3.1t
• Protein Accession #: NP_003484.1
• Purification: Affinity purified
• Nucleotide Accession #: NM_003493.2
• Target Post-Translational Modification: Tri-Methyl-Lys9
• Gene Symbol: HIST3H3
• Predicted Species Reactivity: Human
• Application: CHIP, IF
• Predicted Homology Based on Immunogen Sequence: Yeast
• Image 1: Human Hela cells
  
  ChIP results obtained with the Aviva recombinant antibody directed against H3K9me3r. ChIP assays were performed using human HeLa cells, the Aviva recombinant antibody against H3K9me3 and optimized PCR primer sets for qPCR. ChIP was performed on sheared chromatin from 100,000 and 5,000 cells. Different amounts of the antibody were analysed. A negative control recombinant antibody 1 or 5 ug/IP) was used as negative IP control. QPCR was performed with primers for the heterochromatin marker Sat2 and for the ZNF510 gene, used as positive controls, and for the promoters of the active EIF4A2 and GAPDH genes, used as negative controls. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).
• Image 2: K562 cells
  
  ChIP-seq results obtained with the Aviva recombinant antibody directed against H3K9me3r. ChIP was performed with 1.3 ug of the Aviva antibody against H3K9me3 on sheared chromatin from 4 million K562 cells. The IP'd DNA was analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The sequenced reads were aligned to human genome version 19 using the ELAND algorithm. Figure 2A shows the signal distribution along the long arm of chromosome 19 and a zoomin to an enriched region containing several ZNF repeat genes. Figure 2B shows the enrichment at ZNF510 and Figure C and D show the enrichment at the MEG3 and KCNQ1 imprinted genes.
• Image 3: NIH 3T3 cells
  
  Immunofluorescence using the Aviva recombinant antibody directed against H3K9me3r. NIH 3T3 cells were stained with the Aviva antibody against H3K9me3, left or with the negative control recombinant antibody, right. The bottom panel shows counterstaining of the cells with DAPI. (Hattori T. et al., 2013).