Description
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: 1:100
Immunohistochemistry (Paraffin): 1:300 - 1:1200
Immunofluorescence (Immunocytochemistry): 1:100 - 1:400
Chromatin IP: 1:25
Peptide ELISA (DELFIA): 1:2000
Storage
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at -20°C. Do not aliquot the antibody.
Protocol
Available protocols: Western Blotting, Immunoprecipitation, Immunohistochemistry (Paraffin), Immunofluorescence (Immunocytochemistry), Chromatin IP
Specificity / Sensitivity
Acetylated-Lysine Antibody detects proteins posttranslationally modified by acetylation on the epsilon-amine groups of lysine residues. This antibody recognizes acetylated lysine in a wide range of sequence contexts. This antibody has been demonstrated to recognize acetylated histones, p53, CBP, PCAF and chemically acetylated BSA. This antibody has been shown to react with as little as 0.04 ng of chemically acetylated BSA while not recognizing up to 25 µg of nonacetylated BSA.
Species Reactivity: All Species Expected
Source / Purification
Polyclonal antibodies are produced by immunizing animals with a synthetic acetylated lysine-containing peptide. Antibodies are purified by protein A and peptide affinity chromatography.
Background
Acetylation of lysine, like phosphorylation of serine, threonine or tyrosine, is an important reversible modification controlling protein activity. The conserved amino-terminal domains of the four core histones (H2A, H2B, H3, and H4) contain lysines that are acetylated by histone acetyltransferases (HATs) and deacetylated by histone deacetylases (HDACs) (1). Signaling resulting in acetylation/deacetylation of histones, transcription factors, and other proteins affects a diverse array of cellular processes including chromatin structure and gene activity, cell growth, differentiation, and apoptosis (2-6). Recent proteomic surveys suggest that acetylation of lysine residues may be a widespread and important form of post-translational protein modification that affects thousands of proteins involved in control of cell cycle and metabolism, longevity, actin polymerization, and nuclear transport (7,8). The regulation of protein acetylation status is impaired in cancer and polyglutamine diseases (9), and HDACs have become promising targets for anti-cancer drugs currently in development (10).
