Histone H3 Antibody (Acetyl-Lys56) (OADC00107)

• Product Number: OADC00107
• Name: Histone H3 Antibody (Acetyl-Lys56) (OADC00107)
• Conjugation: Unconjugated
• NCBI Gene Id: 8290; 8350; 8351; 8352; 8353; 8354; 8355; 8356; 8357; 8358; 8968
• Host: Rabbit
• Clonality: Polyclonal
• Concentration: 0.6 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: Polyclonal antibody raised in rabbit against the region of histone H3 containing the acetylated lysine 56 (H3K56ac), using a KLH-conjugated synthetic peptide.
• Application Info: CHIP: 5 ug/CHIP
  ELISA: 1:500
  Dot Blotting: 1:20,000
• Uniprot ID: P68431
• Protein Name: histone H3.1t
• Protein Accession #: NP_003520.1
• Purification: Affinity purified
• Nucleotide Accession #: NM_003529.2
• Target Post-Translational Modification: Acetyl-Lys56
• Preservative: 0.05% Sodium Azide and 0.05% ProClin 300.
• Gene Symbol: HIST3H3
• Predicted Species Reactivity: Human
• Application: CHIP, DB
• Image 1: Human Hela cells
  
  ChIP results obtained with the Aviva antibody directed against H3K56acr. ChIP assays were performed using human HeLa cells, the Aviva antibody against H3K56ac and optimized PCR primer sets for qPCR. ChIP was peformed using sheared chromatin from 1.5 million cells. A titration of the antibody consisting of 0.5, 1, 2 and, 5 ug per ChIP experiment was analysed. IgG (1 ug/IP) was used as negative IP control. QPCR was performed with primers for a region approximately 1 kb upstream of the GAPDH promoter and for the EIF4A2 promoter, used as positive controls, and for the coding region of the inactive MYOD1 gene and the Sat2 satellite repeat, 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: HelaS3 cells
  
  ChIP-seq results obtained with the Aviva antibody directed against H3K56acr. ChIP was performed on sheared chromatin from 1.5 million HeLaS3 cells using 5 ug of the Aviva antibody against H3K56ac as described above. The IP'd DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure shows the enrichment along the complete sequence and a 1 Mb region of the X-chromosome and in genomic regions of chromosome 12 and 3, surrounding the GAPDH and EIF4A2 positive control genes.
• Image 3: H3K56acr
  
  Cross reactivity tests using the Aviva antibody directed against H3K56acr. To test the cross reactivity of the Aviva antibody against H3K56ac, a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K56. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest.
• Image 4: H3K56ac
  
  Determination of the antibody titer. To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Aviva antibody directed against H3K56ac in antigen coated wells. 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 1:15,300.