AAC03-HRP, is an HRP labeled pan-acetyl lysine mouse monoclonal antibody that is part of the Signal-Seeker™ product line. AAC03-HRP anti-acetyl-lysine antibody, clone 19C4B2.1, has been tested in western blot applications and has been shown to be at least 10 times more potent than using unconjugated AAC03 with standard mouse-HRP secondary. The AAC03, anti-acetyl lysine antibody recognizes proteins post-translationally modified by acetylation on the epsilon amine groups of lysine residues that occur on 30-50% of all proteins and in particular histones, p53, tubulin and myosin. A proprietary mixture of acetylated proteins was used to produce the 19C4B2.1, highly robust antibody that has been shown to recognize a wide range of acetylated proteins.
Western Blot using Acetyl-Lysine Antibody (AAC03-HRP)
Fig 1: A: Murine tissue extract, 30 μg brain extract. B: 30 μg of Cos-7 cell lysate treated with TSA and nicotinamide (+) or untreated (-). Strongly enhanced bands at 55 and 14-16 kDa in TSA-treated lysate correspond to acetylated tubulin and histone proteins, respectively. C: Titration of acetylated BSA. Lanes 1-5 contain 0.5, 0.1, 0.05, 0.01, and 0.005 ng Ac-BSA, lanes 6-7 contain 500 and 1000 ng non-acetylated BSA, respectively. AAC03-HRP was used at a 1:3000 dilution following the recommended western blot protocol.
Acetylation of proteins can occur as a co-translational or post-translational modification (PTM) (1). Co-translational acetylation occurs at the N-terminal of approximately 85% of mammalian proteins, it is irreversible and is thought to be important in protein stability, localization and synthesis (1). Post-translational acetylation occurs on the epsilon amino group of lysine residues as a reversible and highly dynamic PTM that is known to be a key regulator in multiple cellular events, including chromatin structure, transcription, metabolism, signal transduction and cytoskeletal regulation (2-3). To date over 4,000 proteins have been identified as targets for PTM acetylation which is comparable to phosphorylation in cellular prevelance (3). Antibody AAC01 detects acetyl lysine PTMs.
1 Bogdan P. and Sherman F. 2002. The diversity of acetylated proteins. Genome Biol. 3 (5): reviews 0006.
2 Lundby A. et al. 2012. Proteomic analysis of lysine acetylation sites in rat tissues reveals organ specificity and cellular patterns. Cell Reports 2:419-431.
3 Sadoul K. et al. 2010. The tale of protein lysine acetylation in the cytoplasm. J. Biomed. Biotech. 2011:1-15.
4 Golemis EA et. Al, Protein-Protein Interactions, CSHLP, 2005, p67