PTMScan® Ubiquitin Remnant Motif (K-ε-GG) Kit #5562
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 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.
Storage
Protocol
Product Description
Background
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: 42
- 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) Apr 15;159-160:146-156.s
- 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.
- Nguyen, D.T.T. et al. (2017) Cell Death Differ 24, 1063-1078.
- Potu, H. et al. (2017) Nat Commun 8, 14449.
- Yi, J.J. et al. (2017) J Biol Chem 292, 12503-12515.
- 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.
- Sohn, S.Y. et al. (2015) J Virol 89, 1744-55.
- Onizawa, M. et al. (2015) Nat Immunol 16, 618-27.
- 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.
- Tong, Z. et al. (2014) Mol Cell Proteomics 13, 2871-82.
- Iesmantavicius, V. et al. (2014) Mol Cell Proteomics 13, 1979-92.
- 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.
- Bustos, D. et al. (2012) Mol Cell Proteomics 11, 1529-40.
- Mathew, R. et al. (2012) Nature 491, 618-21.
- Udeshi, N.D. et al. (2012) Mol Cell Proteomics 11, 148-59.
- Lee, K.A. et al. (2011) J Biol Chem 286, 41530-8.
- Kim, W. et al. (2011) Mol Cell 44, 325-40.
- Emanuele, M.J. et al. (2011) Cell 147, 459-74.
- Zhang, N. et al. (2017) Sci Rep 7, 13601.
Limited Uses
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