Product Usage Information
For optimal ChIP 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
Immunohistochemistry (Paraffin): 1:1600
Immunofluorescence (Immunocytochemistry): 1:800
Flow Cytometry (Fixed/Permeabilized): 1:50
Chromatin IP: 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, Immunohistochemistry (Paraffin), Immunofluorescence (Immunocytochemistry), Flow Cytometry (Fixed/Permeabilized), Chromatin IP
Specificity / Sensitivity
Acetyl-Histone H2B (Lys20) (D7O9W) Rabbit Monoclonal Antibody recognizes endogenous levels of histone H2B protein when acetylated at Lys20. This antibody shows very slight cross-reactivity with histone H2B acetylated at Lys12.
Species Reactivity: Human, Mouse, Rat
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic acetylated peptide corresponding to residues surrounding Lys20 of human histone H2B protein.
Background
The nucleosome, made up of four core histone proteins (H2A, H2B, H3, and H4), is the primary building block of chromatin. Originally thought to function as a static scaffold for DNA packaging, histones have now been shown to be dynamic proteins, undergoing multiple types of post-translational modifications, including acetylation, phosphorylation, methylation, and ubiquitination (1,2). Histone acetylation occurs mainly on the amino-terminal tail domains of histones H2A (Lys5), H2B (Lys5, 12, 15, and 20), H3 (Lys9, 14, 18, 23, 27, 36, and 56), and H4 (Lys5, 8, 12, and 16) and is important for the regulation of histone deposition, transcriptional activation, DNA replication, recombination, and DNA repair (1-3). Hyper-acetylation of the histone tails neutralizes the positive charge of these domains and is believed to weaken histone-DNA and nucleosome-nucleosome interactions, thereby destabilizing chromatin structure and increasing the accessibility of DNA to various DNA-binding proteins (4,5). In addition, acetylation of specific lysine residues creates docking sites for a protein module called the bromodomain, which binds to acetylated lysine residues (6). Many transcription and chromatin regulatory proteins contain bromodomains and may be recruited to gene promoters, in part, through binding of acetylated histone tails. Histone acetylation is mediated by histone acetyltransferases (HATs), such as CBP/p300, GCN5L2, PCAF, and Tip60, which are recruited to genes by DNA-bound protein factors to facilitate transcriptional activation (3). Deacetylation, which is mediated by histone deacetylases (HDAC and sirtuin proteins), reverses the effects of acetylation and generally facilitates transcriptional repression (7,8).
Alternate Names
H2B; H2B clustered histone 3; H2B histone family, member F; H2B-clustered histone 3; H2B.1; H2B/f; H2B1B; H2BC3; H2BFF; HIST1H2BB; histone 1, H2bb; histone cluster 1 H2B family member b; histone cluster 1, H2bb; Histone H2B type 1-B; Histone H2B.1; Histone H2B.f; MGC119804
