PTMScan® Phospho-Akt Substrate Motif mAb 2 (RXRXXS*/T*) Kit #5563
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.
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Protocol
Product Description
Background
- Rush, J. et al. (2005) Nat Biotechnol 23, 94-101.
- Marte, B.M. and Downward, J. (1997) Trends Biochem Sci 22, 355-8.
- Jiang, B.H. et al. (2000) Proc Natl Acad Sci USA 97, 1749-53.
- Scheid, M.P. and Woodgett, J.R. (2000) Curr Biol 10, R191-4.
- Montminy, M. (1997) Annu Rev Biochem 66, 807-22.
- Pearson, R.B. and Kemp, B.E. (1991) Methods Enzymol 200, 62-81.
- Manning, B.D. and Cantley, L.C. (2007) Cell 129, 1261-74.
- Moritz, A. et al. (2010) Sci Signal 3, ra64.
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