Histone H2A (Pan) Antibody (OADC00062)

Catalog No: OADC00062

Size:50ug

Western blot analysis using the Aviva antibody directed against H2Apanr. Western blot was performed on whole cell (25 ug, lane 1) and histone extracts (15 ug, lane 2) from HeLa cells, and on 1 ug of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Aviva antibody against H2Apan. The antibody was diluted 1:2,000 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.

ChIP results obtained with the Aviva antibody directed against H2Apanr. ChIP assays were performed using human HeLa cells, the Aviva antibody against H2Apan and optimized PCR primer sets for qPCR. ChIP was performed with the Auto Histone ChIP-seq" kit on sheared chromatin from 1 million cells using the IP-Star automated system. A titration of the antibody consisting of 1, 2, 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 GAPDH and EIF4A2 promoters, used as negative controls and for the inactive MYOD1 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).

Determination of the titer. To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Aviva antibody directed against H2Apan in antigen coated wells. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:32,500.

Product Info

• Predicted Species Reactivity: Human
• Product Format: Liquid. Provided in PBS containing 0.05% azide and 0.05% ProClin 300.
• Clonality: Polyclonal
• Host: Rabbit
• Conjugation: Unconjugated
• Application: CHIP, WB
• Reconstitution and Storage: Store at -20C. Avoid repeated freeze/thaw cycles.
• Purification: Affinity purified
• Concentration: 4 mg/mL
• Preservative: 0.05% Sodium Azide and 0.05% ProClin 300.
• Specificity: Polyclonal antibody raised in rabbit against histone H2A using a KLH-conjugated synthetic peptide containing an unmodified sequence from the C-terminal part of the protein.
• Application Info: CHIP: 1-2 ug/CHIP
  ELISA: 1:100-1:1,000
  Western Blotting: 1:2,000

Target Info

• Gene Symbol: HIST1H2AE
• Gene Full Name: histone cluster 1, H2ae
• Alias Symbols: H2A.1, H2A.2, H2A/a, H2AC4, H2AFA, HIST1H2AE
• NCBI Gene Id: 3012; 317772; 8335; 8338
• Protein Name: histone H2A type 1-B/E
• 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 H2A 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.
• Uniprot ID: Q7L7L0
• Protein Accession #: NP_254280.1
• Nucleotide Accession #: NM_033445.2