Thermo Scientific™ Tryptic Digestion Kits

Description

• Accurate identification and analysis of proteins is a central component of proteomic strategies for studying cellular functions and processes
• Using mass spectrometry, proteins and relevant post-translational modifications can be accurately identified, but the method is highly dependent on the quality of the sample preparation
• Thermo Scientific Pierce Digestion Kits provide enzymes and reagents for efficient, reproducible digestion and guanidination to improve peptide fragment ionization
• Sample preparation is one of the most variable and time-consuming steps in the analysis of proteins by MS, and the quality and reproducibility of sample extraction and preparation significantly impact the results
• Occasionally, additional steps are required to improve sample quality, including digestion and recovery of peptide fragments from SDS-PAGE gels, or peptide guanidination to improve sequence coverage
• Each kit has been optimized to produce high quality peptide digests for dependable proteomics analysis using mass spectrometry
• Pierce In-Solution Tryptic Digestion and Guanidination Kit The Pierce In-Solution Tryptic Digestion and Guanidination Kit provides an optimized procedure and reagents for approximately 90 digests, each containing 0.025 to 10 μg of protein
• • Complete digestion —trypsin completely digests protein ranging from 0.025 to 10 μg (∼90 reactions) • Specific —guanidination reaction produces little to no N-terminal modification • Convenient —kit includes reagents for digestion, reduction, alkylation, and guanidination • Efficient —protein can be digested and guanidinated in a single day Trypsin specifically cleaves peptide bonds at the carboxyl side of arginine and lysine residues
• The signals from the arginine-containing peptides are generally stronger, due to the more basic side chain
• To enhance overall ionization, guanidination converts lysines to homoarginines and is specific for the amine of lysine, but may occur minimally at the amine of the peptide's N-terminus
• This derivatization leads to an increase in the intensity of the lysine-containing peptides and improved sequence coverage overall
• Pierce In-Gel Tryptic Digestion Kit The Pierce In-Gel Tryptic Digestion Kit provides a complete set of reagents for performing approximately 150 digestions on colloidal Coomassie- or fluorescent dye-stained protein bands
• The kit includes modified porcine trypsin, de-staining buffers, reduction reagents, alkylation reagents, and digestion buffers, along with detailed and simple instructions
• • Complete —includes all necessary reagents for gel destaining, reduction, alkylation, and tryptic digestion of proteins • Robust —optimized procedure and reagents produce reliable digestions and data generation using a wide range of conditions • Specific —contains highly purified and modified MS-grade trypsin with no chymotryptic activity and minimal autolytic activity Two key sample preparation steps are the complete digestion of proteins into peptides and subsequent concentration and clean-up of peptides for mass spectrometry analysis
• The Pierce In-Gel Tryptic Digestion Kit accomplishes the first of these steps for protein samples extracted from gel slices
• Pierce C-18 Spin Columns and Tips accomplish the second step to clean-up the sample to be MS-ready.