Product Pathways - Motif Antibodies

Acetylated-Lysine Antibody #9441

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Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IF-IC=Immunofluorescence (Immunocytochemistry)  E-P=ELISA (Peptide)
Reactivity Key: All=All species expected

Specificity / Sensitivity

Acetylated-Lysine Antibody detects proteins posttranslationally modified by acetylation on the epsilon-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 nonacetylated 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

Polyclonal antibodies are produced by immunizing rabbits with a synthetic acetylated lysine-containing peptide (KLH-coupled). Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western blot analysis of extracts from NIH/3T3 cells, untreated or sodium butyrate-treated (5 mM for 24 hours), showing an increase in histone acetylation using Acetylated-Lysine Antibody.

Western Blotting

Specificity and sensitivity of Acetylated-Lysine Antibody assayed on acetylated BSA (4; 1; 0.2; 0.04 or 0.008 ng in lanes 1-5) or nonacetylated BSA (25,000; 5,000; 1,000 or 200 ng in lanes 6-9).

Western Blotting

Western blot analysis of extracts from COS cells, untreated or TSA-treated, grown in 10% FBS (lanes 1 and 2) or serum starved for 18 hours (lanes 3 and 4), using Acetylated-Lysine Antibody (upper) or p44/42 MAP Kinase Antibody #9102 (lower).

IP

Western blot analysis of immunoprecipitated p53 showing an increase in p53 acetylation using Acetylated-Lysine Antibody (upper) or p53 antibody (lower). p53 was immunoprecipitated from lysates from 293 cells, untreated or UV-treated, using p53 Antibody #9282.

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Acetylated-Lysine Antibody.

IHC-P (paraffin)

Immunohistochemical staining of a paraffin-embedded human breast tumor section showing nuclear and cytoplasmic localization of proteins with acetylated lysine residues using Acetylated-Lysine Antibody.

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded NIH/3T3 untreated (left) or TSA-treated (right) using Acetylated-Lysine Antibody.

IF-IC

Confocal immunofluorescent analysis of NIH/3T3 cells, untreated (left) or SAHA-treated (right), labeled with Acetylated-Lysine Antibody (green). Actin filaments have been labeled with Alexa Fluor R 555 phalloidin (red). Blue pseudocolor = DRAQ5 TM (fluorescent DNA dye).

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). The regulation of protein acetylation status is impaired in the pathologies of cancer and polyglutamine diseases (7), and HDACs have become promising targets for anti-cancer drugs currently in development (8).

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, 786-794.
3. Liu, L. et al. (1999) Mol. Cell. Biol. 19(2), 1202-1209.
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, 269-278.
7. Hughes, R.E. (2002) Curr. Biol. 12, R141-R143.
8. Vigushin, D.M. and Coombes, R.C. (2004) Curr. Cancer Drug Targets 4, 205-218.

Application References

• Chan, H.M. et al. (2001) Acetylation control of the retinoblastoma tumour-suppressor protein. Nat. Cell Biol. 3, 667-674. This article references the use of Acetylated-Lysine Antibody in the following applications: Western Blot
• Martinez-Balbas, M.A. et al. (2000) Regulation of E2F1 activity by acetylation. EMBO J. 19, 662-671. This article references the use of Acetylated-Lysine Antibody in the following applications: Immunoprecipitation
• Yu, X. et al. (2002) Modulation of p53, ErbB1, ErbB2, and Raf-1 expression in lung cancer cells by depsipeptide FR901228. J. Natl. Cancer Inst. 94 (7), 504-513. This article references the use of Acetylated-Lysine Antibody in the following applications: Western Blotting
• Sano, Y. and Ishii, S. (2001) Increased affinity of c-Myb for CREB-binding protein (CBP) after CBP-induced acetylation. J. Biol. Chem. 276, 3674-3682. This article references the use of Acetylated-Lysine Antibody in the following applications: Western Blotting
• Mal, A. et al. (2001) A role for histone deactylase HDAC1 in modulating the transcriptional activity of MyoD: inhibition of the myogenic program. EMBO J. 20, 1739-1753. This article references the use of Acetylated-Lysine Antibody in the following applications: Western Blot
• Li, S. et al. (2000) Regulation of the homeodomain CCAAT displacement/cut protein function by histone acetyltransferases p300/CREB-binding protein (CBP)-associated factor and CBP. Proc. Natl. Acad. Sci USA 97, 7166-7171. This article references the use of Acetylated-Lysine Antibody in the following applications: Western Blot
• Chen, L. et al. (2001) Duration of nuclear NF-kappaB action regulated by reversible acetylation. Science 293, 1653-1657. This article references the use of Acetylated-Lysine Antibody in the following applications: Western Blotting
• Fu, M. et al. (2000) p300 and p300/cAMP-response element-binding protein-associated factor acetylate the androgen receptor at sites governing hormone-dependent transactivation. J. Biol. Chem. 275, 20853-20860. This article references the use of Acetylated-Lysine Antibody in the following applications: IP
• Qiu, Y. et al. (2004) Acetylation of the BETA2 transcription factor by p300-associated factor is important in insulin gene expression. J. Biol. Chem. 279, 9796-9802. This article references the use of Acetylated-Lysine Antibody in the following applications: IP
• Ard, P.G. et al. (2002) Transcriptional regulation of the mdm2 oncogene by p53 requires TRRAP acetyltransferase complexes. Mol. Cell. Biol. 22, 5650-5661. This article references the use of Acetylated-Lysine Antibody in the following applications: IP
• Bereshchenko, O.R. et al. (2002) Acetylation inactivates the transcriptional repressor BCL6. Nat. Genet. 32, 606-613. This article references the use of Acetylated-Lysine Antibody in the following applications: Western Blotting
• Yuan, Z. et al. (2005) Stat3 Dimerization Regulated by Reversible Acetylation of a Single Lysine Residue. Science. 307, 269-273. This article references the use of Acetylated-Lysine Antibody in the following applications: Western Blotting

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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 commerical licensing terms please contact CST Business Development at cbunker@cellsignal.com.