Host/IsotypeMouse / IgG1 Clone19C4B2.1 Species ReactivityAll species Validation DataAc-K Antibody White Paper

AAC03-HRP, is an HRP labeledpan-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 usingAcetyl-Lysine Antibody (AAC03-HRP) Fig 1:A: Murine tissue extract, 30 mg brain extract. B: 30 mg 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. To see the full Western blot comparison, see the Optimized Protocols or theproduct datasheet.

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.

References

1Bogdan P. and Sherman F. 2002. The diversity of acetylated proteins. Genome Biol. 3 (5): reviews 0006.

2Lundby A. et al. 2012. Proteomic analysis of lysine acetylation sites in rat tissues reveals organ specificity and cellular patterns. Cell Reports 2:419-431.

3Sadoul K. et al. 2010. The tale of protein lysine acetylation in the cytoplasm. J. Biomed. Biotech. 2011:1-15.

4Golemis EA et. Al, Protein-Protein Interactions, CSHLP, 2005, p67

For more information contact: signalseeker@cytoskeleton.com

Associated Products:

Signal-Seeker™ Acetyl-Lysine Detection Kit (Cat. # BK163)

Signal-Seeker™: BlastR™ Rapid Lysate Prep Kit (Cat. # BLR01)

Signal-Seeker™ Acetylation Affinity Beads (Cat.# AAC04-beads)

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