Description
Product Usage Information
For optimal ChIP and ChIP-seq results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 x 10 6 cells) per IP. This antibody has been validated using SimpleChIP ® Enzymatic Chromatin IP Kits.
Western Blotting: 1:1000
Immunoprecipitation: 1:200
Flow Cytometry (Fixed/Permeabilized): 1:50
Chromatin IP: 1:50
Chromatin IP-seq: 1:50
Storage
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at -20°C. Do not aliquot the antibody.
Protocol
Available protocols: Western Blotting, Immunoprecipitation, Flow Cytometry (Fixed/Permeabilized), Chromatin IP, Chromatin IP-seq
Specificity / Sensitivity
Acetyl-Histone H2A.Z (Lys4/Lys7) (D3V1I) Rabbit Monoclonal Antibody recognizes endogenous levels of histone H2AZ protein only when acetylated at Lys4 and/or Lys7. This antibody does not cross-react with other acetylated histones, including histone H2A acetylated at Lys5. This antibody also detects a band around 22 kDa, which is most likely monoubiquitylated histone H2AZ that is acetylated on Lys4 and Lys7.
Species Reactivity: Human, Mouse, Rat
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding acetylated Lys4 and Lys7 of human H2AZ protein.
Background
Modulation of chromatin structure plays a critical role in the regulation of transcription in eukaryotes. The nucleosome, made up of four core histone proteins (H2A, H2B, H3 and H4), is the primary building block of chromatin. In addition to the growing number of post-translational histone modifications regulating chromatin structure, cells can also exchange canonical histones with variant histones that can directly or indirectly modulate chromatin structure (1). There are five major variants of histone H2A: canonical H2A (most abundant), H2A.X, MacroH2A, H2ABbd and H2A.Z (2). Histone H2A.Z, the most conserved variant across species, functions as both a positive and negative regulator of transcription and is important for chromosome stability (2). Several homologous protein complexes, such as SWR-C ( ), TIP60 ( ) and SRCAP (mammals), have been shown to catalyze the ATP-dependent exchange of H2A.Z for H2A in the nucleosome (3,4,5). This exchange of histone H2A variants changes histone-histone interactions in the nucleosome core and alters an acidic patch on the surface of the nucleosome, resulting in changes in nucleosome stability and binding of non-histone proteins such as HP1α (6,7). Acetylation of Histone H2AZ correlates with gene activity (8). Acetylation of Histone H2AZ on Lys4 and Lys7 occurs at the 5' end of genes and confers nucleome destabilization and open chromatin confirmation required for tanscriptional activation (9-11).
Alternate Names
H2A histone family member Z; H2A histone family, member Z; H2A.z; H2A.Z variant histone 1; H2A.Z-1; H2A/z; H2AFZ; H2AZ; H2AZ histone; H2AZ1; Histone H2A.Z; MGC117173
