SigmaFASTä Protease Inhibitor Cocktail Tablet, EDTA Free Catalog Number S8830 Storage Temperature 2–8 °C Product Description Specific inhibitory properties of the components are: Crude cell extracts contain a myriad of endogenous enzymes, such as proteases, which are capable of AEBSF – [4-(2-Aminoethyl)benzenesulfonyl fluoride degrading the proteins present in the sample. The best hydrochloride] – serine proteases, e.g., trypsin, way to preserve the integrity of the proteins is to add a chymotrypsin, plasmin, kallikrein, and thrombin broad spectrum of protease inhibitors tailored to the sample and task. Bestatin hydrochloride – aminopeptidases, e.g., leucine aminopeptidase and alanyl aminopeptidase1-4 The SigmaFASTä Protease Inhibitor Cocktail Tablet, EDTA Free is a mixture of protease inhibitors with a Leupeptin – serine and cysteine proteases, e.g., broad specificity for the inhibition of serine, cysteine, trypsin, plasmin, trypsinogen, urokinase, and aspartic, and metalloproteases. The tablets have been kallikrein3 formulated to be compatible with sample processing by immobilized metal affinity chromatography (IMAC) such E-64 – [N-(trans-Epoxysuccinyl)-L-leucine as used for the purification of histidine-tagged proteins. 4-guanidinobutylamide] – cysteine proteases, e.g., The tablets contain no strong metal chelators, such as calpain, papain, cathepsin B, and cathepsin L1 EDTA, which might interfere with IMAC processing. Finally, the tablets contain a unique, proprietary Aprotinin – serine protease, e.g., chymotrypsin, trypsin, formulation including a pepstatin A salt to provide and elastase additional protection not normally obtained in a tablet format. The broad protection afforded allows these Pepstatin A – acid proteases, e.g., pepsin, renin, and tablets to be used for the purification of histidine-tagged cathepsin D, and many microbial aspartic proteins and as a general-purpose protease inhibitor proteases cocktail. Phosphoramidon disodium salt – thermolysin and Components collagenase Each tablet can be used to prepare 100 ml of 1´ protease inhibitor solution, which contains the Precautions and Disclaimer following inhibitor concentrations: This product is for R&D use only, not for drug, household, or other uses. Please consult the Material · AEBSF 2 mM Safety Data Sheet for information regarding hazards · Phosphoramidon 1 mM and safe handling practices. · Bestatin 130 mM · E-64 14 mM Preparation Instructions · Leupeptin 1 mM One tablet generates 100 ml of 1´ protease inhibitor · Aprotinin 0.2 mM solution. Each tablet can be reconstituted in either water or extraction/lysis buffer. · Pepstatin A 10 mM A tablet may also be used to prepare a 10´ protease inhibitor solution, which can be diluted as needed. Concentrations greater than 1´ may appear slightly hazy. This will not affect the performance of the protease inhibitors. Mix the 10´ protease inhibitor solution (15 minutes) until uniformly suspended and dilute as appropriate. Storage/Stability References The tablets are stable as supplied for at least 2 years at 1. Aoyagi, T., and Umezawa, H., Acta Biol. Med. Ger., 2–8 °C. 40, 1523-1529 (1981). 2. Umezawa, H., Ann. Rev. Microbiol., 36, 75-99 The reconstituted protease inhibitor solution (1982). (1´ or 10´) is stable for at least 2 weeks at 2–8 °C. 3. Mumford, R.A., et al., Biochem. Biophys. Res. Comm., 103, 565-572 (1981). Procedure 4. Aoyagi, T., et al, Biochem. Int., 9, 405-411 (1984). One tablet is recommended for the inhibition of the proteases present in a maximum of 20 g of cell mass in SigmaFAST is a registered trademark of Sigma- 100 ml of extract. The inhibitor mixture may be AldrichÒ Biotechnology LP and Sigma-Aldrich Co. dissolved directly in the extraction reagent or added from the 10´ protease inhibitor solution immediately JGD/RMT/MAM 05/09-1 following extraction. Since not all organisms contain the same amount of endogenous proteases it may sometimes be necessary to increase the concentration of inhibitors. Sigma brand products are sold through Sigma-Aldrich, Inc. Sigma-Aldrich, Inc. warrants that its products conform to the information contained in this and other Sigma-Aldrich publications. Purchaser must determine the suitability of the product(s) for their particular use. Additional terms and conditions may apply. Please see reverse side of the invoice or packing slip..