PTMScan® Pilot Ubiquitin Remnant Motif (K-ε-GG) Kit #14482
Motif analysis was done using all ubiquitin remnant peptides in a ubiquitin remnant motif antibody PTMScan® study. Cellular tryptic peptides from HCT 116 cells, untreated or treated with MG-132 #2194, were immunoprecipitated with ubiquitin remnant motif antibody K-ε-GG and analyzed by OrbiTrap™ LC-MS. The study gave 599 non-redundant sites. The motif logo shows that the K-ε-GG antibody is a general motif antibody that recognizes the K-ε-GG motif and does not have other amino acid preferences.
To Purchase # 14482
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® Ubiquitin Branch Motif (K-ε-GG) Immunoaffinity Beads #1990 | 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 Ubiquitin Remnant Motif (K-ε -GG) Kit has a higher sensitivity and specificity magnetic bead version: PTMScan® HS Ubiquitin/SUMO Remnant Motif (K-ε -GG) Kit in 10-assay (#59322) or 3-assay (#19089) formats, as well as a bead-only (#34608) format.
PTMScan® Pilot Ubiquitin Remnant Motif (K-ε -GG) Kit has a higher sensitivity and specificity magnetic bead version: PTMScan® HS Ubiquitin/SUMO Remnant Motif (K-ε -GG) Kit in 10-assay (#59322) or 3-assay (#19089) formats, as well as a bead-only (#34608) format.
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
Ubiquitin is a conserved polypeptide unit that plays an important role in the ubiquitin-proteasome pathway. Ubiquitin can be covalently linked to many cellular proteins by the ubiquitination process, which targets proteins for degradation by the 26S proteasome. Three components are involved in the target protein-ubiquitin conjugation process. Ubiquitin is first activated by forming a thiolester complex with the activation component E1; the activated ubiquitin is subsequently transferred to the ubiquitin-carrier protein E2, then from E2 to ubiquitin ligase E3 for final delivery to the epsilon-NH2 of the target protein lysine residue (2-4). The ubiquitin-proteasome pathway has been implicated in a wide range of normal biological processes and in disease-related abnormalities. Several proteins such as IκB, p53, cdc25a and bcl-2 have been shown to be targets for the ubiquitin-proteasome process as part of regulation of cell cycle progression, differentiation, cell stress response, and apoptosis (5-8).In this assay, the PTMScan® Ubiquitin Remnant Motif (K-ε-GG) Antibody Bead Conjugate, a proprietary ubiquitin branch (“K-ε-GG”) antibody with specificity for a di-glycine tag that is the remnant of ubiquitin left on protein substrates after trypsin digestion, is used to enrich ubiquitinated peptides from trypsin digested cell samples. This enrichment is followed by LC-MS/MS analysis for quantitative profiles of hundreds to over a thousand non-redundant ubiquitinated sequences.
- Rush, J. et al. (2005) Nat Biotechnol 23, 94-101.
- Ciechanover, A. (1998) EMBO J 17, 7151-60.
- Hochstrasser, M. (2000) Nat Cell Biol 2, E153-7.
- Hochstrasser, M. (2000) Science 289, 563-4.
- Bernardi, R. et al. (2000) Oncogene 19, 2447-54.
- Aberle, H. et al. (1997) EMBO J 16, 3797-804.
- Salomoni, P. and Pandolfi, P.P. (2002) Nat Cell Biol 4, E152-3.
- Jesenberger, V. and Jentsch, S. (2002) Nat Rev Mol Cell Biol 3, 112-21.
Product Citations: 41
- McCann, J.J. et al. (2020) Cancer Res 80, 430-443.
- Yasuda, S. et al. (2020) Nature 578, 296-300.
- Tufegdzic Vidakovic, A. et al. (2019) Methods 159-160, 146-156.
- Zeng, F. et al. (2019) PLoS Biol 17, e3000525.
- Chen, X.L. et al. (2018) Front Plant Sci 9, 1064.
- Bakos, G. et al. (2018) Nat Commun 9, 4776.
- Christensen, D.G. et al. (2018) MBio 9.
- Wade, B.E. et al. Pulm Circ 8, 2045894018788267.
- Brockmann, M. et al. (2017) Nature 546, 307-311.
- Dogan, T. et al. (2017) J Biol Chem 292, 14311-14324.
- Lumpkin, R.J. et al. (2017) Nat Commun 8, 1171.
- Yi, J.J. et al. (2017) J Biol Chem 292, 12503-12515.
- Potu, H. et al. (2017) Nat Commun 8, 14449.
- Nguyen, D.T.T. et al. (2017) Cell Death Differ 24, 1063-1078.
- de Almagro, M.C. et al. (2017) Cell Death Differ 24, 26-37.
- Huebner, A.R. et al. (2016) Mol Cell Proteomics 15, 1556-71.
- Ivanova, E. and Carpino, N. (2016) Mol Immunol 73, 19-28.
- Li, X.M. et al. (2015) Nat Genet 47, 827-33.
- Onizawa, M. et al. (2015) Nat Immunol 16, 618-27.
- Sohn, S.Y. et al. (2015) J Virol 89, 1744-55.
- Varfolomeev, E. et al. (2015) Biochem J 466, 45-54.
- Yoshida, Y. et al. (2015) Proc Natl Acad Sci U S A 112, 4630-5.
- Yu, K. et al. (2015) J Mol Biol 427, 2121-34.
- Ryder, D.J. et al. (2015) PLoS One 10, e0136247.
- Gispert, S. et al. (2015) Hum Mol Genet 24, 1061-76.
- Krönke, J. et al. (2014) Science 343, 301-5.
- Theurillat, J.P. et al. (2014) Science 346, 85-89.
- Thompson, J.W. et al. (2014) J Biol Chem 289, 28942-55.
- Iesmantavicius, V. et al. (2014) Mol Cell Proteomics 13, 1979-92.
- Tong, Z. et al. (2014) Mol Cell Proteomics 13, 2871-82.
- Tammsalu, T. et al. (2014) Sci Signal 7, rs2.
- Anania, V.G. et al. (2014) Mol Cell Proteomics 13, 145-56.
- Udeshi, N.D. et al. (2013) Nat Protoc 8, 1950-60.
- Carrano, A.C. and Bennett, E.J. (2013) Mol Cell Proteomics 12, 3521-31.
- Buckley, S.M. et al. (2012) Cell Stem Cell 11, 783-98.
- Mathew, R. et al. (2012) Nature 491, 618-21.
- Udeshi, N.D. et al. (2012) Mol Cell Proteomics 11, 148-59.
- Bustos, D. et al. (2012) Mol Cell Proteomics 11, 1529-40.
- Emanuele, M.J. et al. (2011) Cell 147, 459-74.
- Lee, K.A. et al. (2011) J Biol Chem 286, 41530-8.
- Kim, W. et al. (2011) Mol Cell 44, 325-40.
限制使用
除非 CST 的合法授书代表以书面形式书行明确同意，否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件，除非书 CST 的合法授书代表以书面形式书独接受， 否书均被拒书，并且无效。
专品专有“专供研究使用”的专专或专似的专专声明， 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的， 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专，且专用于研专用途。将专品用于专断、专防或治专目的， 或专专售（专独或作专专成）或其他商专目的而专专专品，均需要 CST 的专独专可。客专：(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品，或使用专品制造任何商专专品，(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专，或使用专品开专任何与 CST 的专品或服专专争的专品或服专， (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专，(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品， (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专
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.
UbiScan is a registered trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit our
Trademark Information page.