Histone H3 Antibody (Tri-Methyl-Lys9) (OADC00272)

• Product Number: OADC00272
• Name: Histone H3 Antibody (Tri-Methyl-Lys9) (OADC00272)
• Isotype: IgG1
• Conjugation: Unconjugated
• NCBI Gene Id: 8290; 8350; 8351; 8352; 8353; 8354; 8355; 8356; 8357; 8358; 8968
• Host: Mouse
• Clonality: Monoclonal
• Concentration: 1 mg/mL
• Gene Full Name: histone cluster 3, H3
• Alias Symbols: H3t, H3.4, H3/g, H3FT, HIST3H3
• Product Format: Liquid. Provided in PBS containing 0.05% azide.
• 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: Monoclonal antibody raised in mouse against histone H3, trimethylated at lysine 9 (H3K9me3), using a KLH-conjugated synthetic peptide. 
• Application Info: CHIP: 1-3 ug/CHIP
  ELISA: 1:1,000
  Western Blotting: 1:1,000
  IF: 1:500
• Uniprot ID: Q16695
• Protein Name: histone H3.1t
• Protein Accession #: NP_003484.1
• Purification: Protein A/G purified
• Nucleotide Accession #: NM_003493.2
• Target Post-Translational Modification: Tri-Methyl-Lys9
• Preservative: 0.05% Sodium Azide
• Gene Symbol: HIST3H3
• Predicted Species Reactivity: Human
• Application: CHIP, WB, IF
• Predicted Homology Based on Immunogen Sequence: fungi
• Image 1: Human Hela cells
  
  ChIP results obtained with the Aviva monoclonal antibody directed against H3K9me3. A. ChIP was performed using HeLa cells, the monoclonal antibody against H3K9me3 and optimized PCR primer sets for qPCR. ChIP was performed using sheared chromatin from 1.6 million cells. A titration of the antibody consisting of 1, 3 and 9 ug per ChIP experiment was analysed. IgG (5 ug/IP) was used as negative IP control. QPCR was performed with primers for the promoter and the coding region of the GAPDH gene, and for the RPL10 and HBB promoters. B. ChIP was performed using sheared chromatin from 1 million K562 cells. A titration of the antibody consisting of 1, 2 and 5 ug per ChIP experiment was analysed. IgG (2 ug/IP) was used as negative IP control. QPCR was performed with primers for the promoters of the GAPDH and EIF4A2 genes, used as negative controls, and for the ZNF12 gene and the Sat2 satellite repeat, used as positive 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: H3K9me3
  
  Cross reactivity of the Aviva monoclonal antibody directed against H3K9me3. To test the specificity an ELISA was performed using a serial dilution of the Aviva monoclonal antibody against H3K9me3. The wells were coated with peptides containing the unmodified H3K9 as well as the mono-, di- and trimethylated H3K9 and the trimethylated H3K4 and H3K27. Figure shows a high specificity of the antibody for the modification of interest.
• Image 3: Hela cells
  
  Immunofluorescence using the Aviva monoclonal antibody directed against H3K9me3. HeLa cells were stained with the Aviva antibody against H3K9me3 and with DAPI. Cells were fixed with 4% formaldehyde for 10' and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H3K9me3 antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.
• Image 4: Human Hela cells
  
  Western blot analysis using the Aviva monoclonal antibody directed against H3K9me3. Histone extracts (15 ug) from HeLa cells were analysed by Western blot using the Aviva monoclonal antibody against H3K9me3 diluted 1:1,000 in TBS-Tween containing 5% skim milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.