PTMScan® Pilot Acetyl-Lysine Motif [Ac-K] Kit #14499
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The chart shows the relative category distribution of proteins with acetylated lysine residues derived from peptides identified from an AcetylScan® LC-MS/MS experiment of mouse liver tissue using PTMScan® Acetyl-Lysine Motif [Ac-K] Immunoaffinity Beads.
To Purchase # 14499
Additional Information
This product is intended for peptide enrichment and mass spectrometry analysis. To learn more about our Proteomics Kits and Services please answer a few questions for our Proteomics group.
- Product Includes
- Related Products
| Product Includes | Volume (with Count) |
|---|---|
| PTMScan® Acetyl-Lysine Motif [Ac-K] Immunoaffinity Beads #13362 | 3 x 80 µl |
| PTMScan® IAP Buffer (10X) #9993 | 3 x 600 µl |
Product Information
Product Usage Information
Cells are lysed in a urea-containing buffer, cellular proteins are digested by proteases, and the resulting peptides are purified by reversed-phase solid-phase extraction. Peptides are then subjected to immunoaffinity purification using a PTMScan® Motif Antibody conjugated to protein A agarose beads. Unbound peptides are removed through washing, and the captured PTM-containing peptides are eluted with dilute acid. Reversed-phase purification is performed on microtips to desalt and separate peptides from antibody prior to concentrating the enriched peptides for LC-MS/MS analysis. CST recommends the use of PTMScan® IAP Buffer #9993 included in the kit.
PTMScan® Pilot Acetyl-Lysine Motif (Ac-K) Kit has a higher sensitivity and specificity magnetic bead version: PTMScan® HS Acetyl-Lysine (Ac-K) Kit in 10-assay (#46784) or 3-assay (#50071) formats.
PTMScan® Pilot Acetyl-Lysine Motif (Ac-K) Kit has a higher sensitivity and specificity magnetic bead version: PTMScan® HS Acetyl-Lysine (Ac-K) Kit in 10-assay (#46784) or 3-assay (#50071) formats.
Storage
Antibody beads are supplied in IAP buffer containing 50% glycerol. Store at -20°C. Do not aliquot the antibody.
Protocol
Product Description
PTMScan® Technology employs a proprietary methodology from Cell Signaling Technology (CST) for peptide enrichment by immunoprecipitation using a specific bead-conjugated antibody in conjunction with liquid chromatography (LC) tandem mass spectrometry (MS/MS) for quantitative profiling of post-translational modification (PTM) sites in cellular proteins. These include phosphorylation (PhosphoScan®), ubiquitination (UbiScan®), acetylation (AcetylScan®), and methylation (MethylScan®), among others. PTMScan® Technology enables researchers to isolate, identify, and quantitate large numbers of post-translationally modified cellular peptides with a high degree of specificity and sensitivity, providing a global overview of PTMs in cell and tissue samples without preconceived biases about where these modified sites occur (1). For more information on PTMScan® Proteomics Services, please visit www.cellsignal.com/services/index.html.
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).
- Hassig, C.A. and Schreiber, S.L. (1997) Curr Opin Chem Biol 1, 300-8.
- Allfrey, V.G. et al. (1964) Proc Natl Acad Sci USA 51, 786-94.
- Liu, L. et al. (1999) Mol Cell Biol 19, 1202-9.
- Boyes, J. et al. (1998) Nature 396, 594-8.
- Polevoda, B. and Sherman, F. (2002) Genome Biol 3, reviews 0006.
- Yoshida, M. et al. (2003) Prog Cell Cycle Res 5, 269-78.
- Kim, S.C. et al. (2006) Mol Cell 23, 607-18.
- Choudhary, C. et al. (2009) Science 325, 834-40.
- Hughes, R.E. (2002) Curr Biol 12, R141-3.
- Vigushin, D.M. and Coombes, R.C. (2004) Curr Cancer Drug Targets 4, 205-18.
Product Citations: 1
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For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
AcetylScan is a registered trademark of Cell Signaling Technology, Inc.
MethylScan is a registered trademark of Cell Signaling Technology, Inc.
PhosphoScan is a registered trademark of Cell Signaling Technology, Inc.
PhosphoSitePlus is a registered trademark of Cell Signaling Technology, Inc.
UbiScan is a registered trademark of Cell Signaling Technology, Inc.
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