PTMScan® Trypsin, TPCK-Treated #56296
Product Information
Product Usage Information
PTMScan® Trypsin-TPCK is provided for use with Cell Signaling Technology's patented PTMScan® protocol in the initial protein digestion step. Samples in urea lysis buffer need to be diluted to 2M or less urea prior to addition of trypsin. For a 1 mg/mL solution, dissolve 20 mg trypsin in 4 mL of 1 mM HCl prior to use. Transfer the 4 mL to a 50 mL conical tube and bring the volume up to 20 mL with 1 mM HCl. Aliquot and store at -80ºC. Consult the specific PTMScan® kit and protocol for more details on the appropriate protease before digesting any protein samples.
Storage
Store lyophilized trypsin powder at 4ºC protected from moisture. Once reconstituted, store trypsin solutions at -80ºC. Lyophilized trypsin has a shelf life of 1 year at 4º C and solutions are stable for 6 months at -80º C.
Product Description
Trypsin is a serine endopeptidase derived from its inactive pancreatic zymogen, trypsinogen, when the N-terminal 6 amino acid leader sequence is enzymatically removed. Activated trypsin cleaves amide and ester bonds of lysine and arginine and is used extensively in detaching cells from culture dishes, protein sequencing, and proteomics applications.
Specificity / Sensitivity
One unit of trypsin hydrolyzes 1 μmole of p-toluene-sulfonyl-L-arginine methyl ester (TAME) per minute at 25ºC, pH 8.0 in the presence of 10 mM calcium. One mg of trypsin is approximately equivalent to 180 TAME units.
Source / Purification
Trypsin is chromatographically purified from bovine pancreas, treated with L-(tosylamido-2-phenyl) ethyl chloromethyl ketone (TPCK) to inhibit chymotryptic activity, 0.22 micron diafiltered against 1 mM HCl, and lyophilized.
Background
Trypsin digests polypeptides by hydrolysis at the carboxyl side of unmodified arginine and lysine residues (1). Proteolysis is slower when the cleavage site is flanked by acidic residues and will not occur if the lysine or arginine is followed by a proline (2-5). Trypsin's activity is optimal at pH 8.0 and is inhibited to varying degrees by organophosphorus compounds such as diisopropyl fluorophosphate, as well as EDTA, aprotinin, Ag+, and benzamidine (6-8).
- Brown, W.E. and Wold, F. (1973) Biochemistry 12, 828-34.
- Rodriguez, J. et al. (2008) J Proteome Res 7, 300-5.
- Leiros, H.K. et al. (2004) Protein Sci 13, 1056-70.
- Rawlings, N.D. and Barrett, A.J. (1994) Methods Enzymol 244, 19-61.
- Perona, J.J. and Craik, C.S. (1995) Protein Sci 4, 337-60.
- Kostka, V. and Carpenter, F.H. (1964) J Biol Chem 239, 1799-803.
- Levilliers, N. et al. (1970) Arch Biochem Biophys 140, 474-83.
- Polgár, L. (2005) Cell Mol Life Sci 62, 2161-72.
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.
PTMScan is a registered trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit our
Trademark Information page.