Cell Signaling Immobilized Trypsin Product Immobilized Trypsin For Research Use Only. Not for Use in Diagnostic Procedures. Description: Immobilized Trypsin provides a fast and convenient method for digesting a range of concentrations of purified protein or complex protein mixtures. Digested peptides are easily separated from the Immobilized Trypsin as they flow through the spin column into the collection tube. Immobilized Trypsin is easily removed from the peptide solution because the trypsin does not pass though the column frit. Trypsin is a proteolytic enzyme, which cleaves at the carboxyl side of positively charged Lysine (Lys) and Arginine (Arg). When these amino acids are followed by the nonpolar Proline (Pro), the digestion of the site is not efficient. When Lys and Arg are followed by acids [Aspartic Acid (Asp) and Glutamic Acid (Glu)] the digestion is also not as efficient. Features: • Fast: Digestions can be accomplished in as little as 30 minutes. • Scalable: Easily adjustable protocol accommodates various protein concentrations. • Easy-to-Use: No shaking or water baths necessary. Storage Conditions: Store at 4°C. Protocol Immobilized Trypsin Technical Manual Chymotrypsin, Sequencing Grade Product Chymotrypsin, Sequencing Grade For Research Use Only. Not for Use in Diagnostic Procedures. Description: Chymotrypsin is a highly-purified serine endopeptidase derived from bovine pancreas that preferentially hydrolyzes at the carboxyl side of aromatic amino acids: Tyr, Phe and Trp. Cleavage may also be observed, but at a lower rate, at Leu and Met. Chymotrypsin activity is optimal in the pH range of 7.0–9.0. This sequencing grade enzyme can be used alone or in combination with other proteases to produce protein digests for peptide mapping applications or protein identification by peptide mass fingerprinting or MS/MS spectral matching. It is suitable for digestion reactions in-solution or in-gel. Storage Conditions: Store at 4°C. Protocol Chymotrypsin Sequencing Grade Product Information Part# 9PIV106 Size Cat.# Price (Fr) 25 µg V1061 82.00 100 µg V1062 263.00 Part# TM077 Size Cat.# Price (Fr) 2 ml V9012 310.00 4 ml V9013 589.00 Trypsin Gold, Mass Spectrometry Grade Product Trypsin Gold, Mass Spectrometry Grade For Research Use Only. Not for Use in Diagnostic Procedures. Description: Trypsin is a serine protease that specifically cleaves at the carboxylic side of lysine and arginine residues. The stringent specificity of trypsin is essential for protein identification. Native trypsin is subject to autolysis, generating pseudotrypsin, which exhibits a broadened specificity including a chymotrypsin-like activity. Such autolysis products, present in a trypsin preparation, would result in additional peptide fragments that could interfere with database analysis of the mass of fragments detected by mass spectrometry. Trypsin Gold, Mass Spectrometry Grade, has been manufactured to provide maximum specificity. Lysine residues in the porcine trypsin have been modified by reductive methylation, yielding a highly active and stable molecule that is extremely resistant to autolytic digestion. The specificity of the purified trypsin is further improved by TPCK treatment, which inactivates chymotrypsin. The treated trypsin is then purified by affinity chromatography and lyophilized to yield Trypsin Gold, Mass Spectrometry Grade. Trypsin is often used for in-gel digestion. The digestion products are purified and concentrated, then analyzed by mass spectrometry to determine their molecular weights. Database searches can then be performed, using the mass of the peptides to identify the protein(s) resolved on the gel. Each lot of quality-tested Trypsin Gold, Mass Spectrometry Grade, is qualified for use with in-gel digestion and mass spectrometric analysis. Features: • Pure: TPCK treatment followed by affinity purification. • Application Qualified: Each lot is qualified by mass spectrometry. • Convenient: Provided in one vial. • Good Value: Stability ensured up to five freeze-thaw cycles, thus minimizing leftover reagents. • Choose Your Configuration: Learn more about our custom options for this product at: www.promega.com/myway/ Storage Conditions: Store the lyophilized powder at –20°C. Reconstitute powder in 50mM acetic acid and store at –20°C. For long-term storage, freeze reconstituted trypsin at –70°C. Limit the number of freeze-thaw cycles to five. Protocol Trypsin Gold, Mass Spectrometry Grade Technical Bulletin Part# TB309 Size Cat.# Price (Fr) 100 µg V5280 194.00 13 For complete and up-to-date product information visit: www.promega.com/catalog 233 Protein Expression and Analysis Pagina 236
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