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Pepsin, Certified MS Grade Catalog Number: PE-103 PRODUCT DESCRIPTION: Pepsin is a digestive protease that is specific for the cleavage at the carboxy terminus side of tyrosine (Y), tryptophan (W), phenylalanine (F), and leucine (L) bonds. Pepsin typically does not cleave at bonds containing valine, alanine, or glycine. Protea s pepsin is highly purified, resulting in a product suitable for proteomics work. Pepsin is an acidic protease, and its inactive zymogen precursor, pepsinogen, is produced in the stomach mucosa. Pepsin digestions are optimally carried out at pH 1 to 3. PROTOCOL: Master stock solution: • Reconstitute the lyophilized pepsin with 100 µL of 10 mM HCl to make a final concentration of 100 ng/µL. • If only a small amount of pepsin is required for the experiment, prepare small aliquots and immediately freeze and store the unused aliquots of pepsin at -20°C to -80°C. In-Solution Digest Working stock solution: • Prepare the Working stock solution immediately before use. • Dilute the Master stock solution (above) to a concentration of 2 ng/µL with 10 mM HCl. Depending on the amount of protein in the samples to be digested, this Working stock concentration can be adjusted. • Any unused portion of the Working stock solution should be immediately frozen at -20°C to -80°C Digestion: • If possible, prepare the protein sample in 10 mM HCl to ensure digestion takes place at pH 3 or lower. • Pepsin should be added to the protein sample at a ratio of 1:50 enzyme to protein by mass. For example, if a sample contains 5 µg of protein, then add 50 µL of the 2 ng/µL pepsin (Working stock solution prepared above in Step B). • Digestion should occur at pH 3 or lower. • Digest for 6 hours to overnight at 37°C. PRODUCT CARE AND STORAGE: Lyophilized enzyme can be stored at -20°C or -80°C for up to 3 years. Reconstituted enzyme can be stored at -20°C for up to 1 month. SPECIFICATIONS: - Lyophilized powder (contains enzyme stabilizers) - Source: porcine stomach - This product is for RandD use only, not for drug, household, or other uses. ORDERING INFORMATION: Catalog Number: PE-103-ea 1 pack of 5 vials, 10 µg/vial (50 µg total) www.proteabio.com ¦ 877.776.8321 ¦ 955 Hartman Run Road, Morgantown, WV 26507