Cell Signaling Technology

Product Pathways - Motif Antibodies

Acetylated-Lysine (Ac-K2-100) Rabbit mAb #9814

Applications Reactivity Sensitivity Isotype
W IP ChIP E-P All Endogenous Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  ChIP=Chromatin IP  E-P=ELISA (Peptide)
Reactivity Key: All=All species expected
Species cross-reactivity is determined by western blot.

Protocols

Specificity / Sensitivity

Acetylated-Lysine (Ac-K2-100) Rabbit mAb detects proteins post-translationally modified by acetylation on the ε-amine groups of lysine residues. The antibody recognizes acetylated lysine in a wide range of sequence contexts. It has been demonstrated to recognize acetylated histones, p53, CBP, PCAF and chemically acetylated BSA. The 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 non-acetylated BSA. (U.S. Patent No's.: 6,441,140; 6,982,318; 7,259,022; 7,344,714; U.S.S.N. 11,484,485; and all foreign equivalents.)

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic acetyl-lysine peptide library. The antibody is formulated from two rabbit monoclones in order to cover broad range of reactivity.

Western Blotting

Western Blotting

Western blot analysis of extracts from COS and HeLa cells, untreated or TSA-treated (1 µM, 6 hours), using Acetylated-Lysine (Ac-K2-100) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from various mouse tissues using Acetylated-Lysine (Ac-K2-100) Rabbit mAb.

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HeLa cells and either 10 μl of Acetylated-Lysine (Ac-K2-100) Rabbit mAb #9814 or 2 μl of Normal Rabbit IgG #2729, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human GAPDH Exon 1 Primers #5516, SimpleChIP® Human RPL30 Exon 3 Primers #7014, SimpleChIP® Human MyoD1 Exon 1 Primers #4490, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.


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 posttranslational 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).

  1. Hassig, C.A. and Schreiber, S.L. (1997) Curr Opin Chem Biol 1, 300-8.
  2. Allfrey, V.G. et al. (1964) Proc Natl Acad Sci USA 51, 786-94.
  3. Liu, L. et al. (1999) Mol Cell Biol 19, 1202-9.
  4. Boyes, J. et al. (1998) Nature 396, 594-8.
  5. Polevoda, B. and Sherman, F. (2002) Genome Biol 3, reviews0006.
  6. Yoshida, M. et al. (2003) Prog Cell Cycle Res 5, 269-78.
  7. Kim, S.C. et al. (2006) Mol Cell 23, 607-18.
  8. Choudhary, C. et al. (2009) Science 325, 834-40.
  9. Hughes, R.E. (2002) Curr Biol 12, R141-3.
  10. Vigushin, D.M. and Coombes, R.C. (2004) Curr Cancer Drug Targets 4, 205-18.

Application References

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Companion Products

Rabbit monoclonal antibody is produced under license (granting certain rights including those under U. S. Patent No. 5,675,063 and 7,429,487) from Epitomics, Inc.License/Use Restrictions: Use of CST Motif Antibodies within certain methods (e.g., U.S. Patent No.'s 7,198,896 & 7,300,753) may require a license from CST. For information regarding academic licensing terms please have your technology transfer office contact CST Legal Department at CST_ip@cellsignal.com. For information regarding commercial licensing terms please contact CST Pharma Services Department at ptmscan@cellsignal.com.

This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

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