Histone H3 Antibody (Methyl-Lys4) (OADC00095)

• Product Number: OADC00095
• Name: Histone H3 Antibody (Methyl-Lys4) (OADC00095)
• Isotype: IgG2b
• 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 and 0.05% ProClin 300.
• 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, monomethylated at lysine 4 (H3K4me1), using a KLH-conjugated synthetic peptide.
• Application Info: CHIP: 1 ug/CHIP
  ELISA: 1:5,000
  Western Blotting: 1:1,000
  IF: 1:500
• Uniprot ID: P68431
• Protein Name: histone H3.1t
• Protein Accession #: NP_003520.1
• Purification: Protein A purified
• Nucleotide Accession #: NM_003529.2
• Target Post-Translational Modification: Methyl-Lys4
• Preservative: 0.05% Sodium Azide and 0.05% ProClin 300.
• Gene Symbol: HIST3H3
• Predicted Species Reactivity: Human
• Application: CHIP, WB, IF
• Image 1: Human Hela cells
  
  ChIP results obtained with the Aviva monoclonal antibody directed against H3K4me1. ChIP assays were performed using HeLa cells, the monoclonal antibody against H3K4me1 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, 5 and 10 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. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).
• Image 2: H3K4me1
  
  Cross reactivity of the Aviva monoclonal antibody directed against H3K4me1. To test the specificity an ELISA was performed using a serial dilution of the Aviva monoclonal antibody against H3K4me1. The wells were coated with peptides containing the unmodified H3K4 as well as the mono-, di- and trimethylated H3K4 and the monomethylated H3K9. 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 H3K4me1. HeLa cells were stained with the Aviva antibody against H3K4me1 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 H3K4me1 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 directed antibody against H3K4me1. Histone extracts (15 ug) from HeLa cells were analysed by Western blot using the Aviva monoclonal antibody against H3K4me1 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.