| Product Description | Aladdin's Protease and Phosphatase Inhibitor Cocktail for Mammalian Cell and Tissue Extracts (50X) includes individually packaged inhibitors for protease and phosphatase, respectively, which can be used for protein extraction from cells or tissues. The Protease Inhibitor Cocktail (50X) consists of 52.5mM AEBSF, 40μM Aprotinin, 2.5mM Bestatin, 0.75mM E64, 1mM Leupeptin and 0.75mM Pepstatin A in high-purity DMSO. The Phosphatase Inhibitor Cocktail (50X) is composed of 250mM sodium fluoride, 50mM sodium pyrophosphate, 50mM β-glycerophosphate and 50mM sodium orthovanadate in H2O. This product also provides one tube of 0.25M EDTA for convenient use.Many endogenous proteases and phosphatases are commonly present in mammalian cell or tissue extracts, which easily leads to degradation or dephosphorylation of protein samples. Therefore, adding appropriate inhibitors is generally necessary for protein extraction..The protease and phosphatase inhibitors in this product are optimized for routine protein extraction from mammalian cells or tissues. It contains serine, cysteine and acidic protease inhibitors with broad inhibitory activity, aminopeptidase inhibitors, as well as serine/threonine, tyrosine, acidic and alkaline phosphatase inhibitors.Different types of protein extracts contain different types of proteases. For example, there is high abundance of serine proteases in mammalian cell extracts, more serine proteases and metalloproteases inbacterial cell extract, and serine, cysteine and metalloproteases in mammalian tissue extracts. Plant extracts often contain large amounts of serine and cysteine proteases, and some contain aspartic proteases. Therefore, Beyotime developed a variety of inhibitor cocktails for the protein extraction from different types of samples. Please refer to Table 1 in appendix for details.This product can effectively inhibit various proteases (e.g., serine protease, aminopeptidase, cysteine protease, threonine and aspartic protease, metalloproteinase) and phosphatases (e.g., serine/threonine, tyrosine, acidic and alkaline phosphatase) in mammalian cell or tissue extracts. Among its components, AEBSF is a water-soluble irreversible inhibitor of serine protease, with an inhibition constant similar to that of PMSF and DFP. It can effectively inhibit trypsin, chymotrypsin, plasmin, kallikrein, thrombin, and other proteases. As a substitute for PMSF and DFP, AEBSF has lower toxicity, better water solubility and better aqueous stability. Aprotinin is a competitive reversible inhibitor of serine protease. Bestatin is a reversible inhibitor of aminopeptidase. E64 is an irreversible inhibitor of cysteine protease. Leupeptin is a reversible inhibitor of serine and cysteine protease. Pepstatin A is a reversible inhibitor of aspartic protease. EDTA, as a chelating agent, is a reversible inhibitor of metalloproteinase. Sodium fluoride is a reversible inhibitor of acid phosphatase. Sodium pyrophosphate is an irreversible inhibitor of serine/threonine phosphatase. β-glycerophosphate is a reversible inhibitor of serine/threonine phosphatase. Sodium orthovanadate is a reversible inhibitor of alkaline phosphatase and tyrosine phosphatase. Please refer to Table 2 in Appendix for specific information of various inhibitors.This product is usually sufficient for preparing 100ml of lysis buffer.
Precautions： It is recommended to store this product in aliquot. This product should be added into solutions immediately before use.This product provides individually packaged EDTA, which can be used accordingly. For example, when used for metalloproteinase activity assays, EDTA should not be added.Protease and phosphatase inhibitors contained in this product are harmful to human. Please take effective measures to avoid direct contact or inhalation.This product is for R&D only. Not for drug, household, or other uses.For your safety and health, please wear a lab coat and disposable gloves during the operation.Appendix：Table 1. Inhibitor cocktails from .Cat. No.Product NamePack SizeP1005Protease Inhibitor Cocktail for General Use, 100X1mlP1006Protease Inhibitor Cocktail for General Use, 100X5mlP1008Protease inhibitor cocktail for general use, MS-SAFE, 50X1mlP1009Protease inhibitor cocktail for general use, MS-SAFE, 50X5mlP1010Protease inhibitor cocktail for mammalian cell and tissue extracts, 100X1mlP1011Protease inhibitor cocktail for mammalian cell and tissue extracts, 100X5mlP1015Protease inhibitor cocktail for plant cell and tissue extracts, 100X1mlP1016Protease inhibitor cocktail for plant cell and tissue extracts, 100X5mlP1020Protease inhibitor cocktail for fungal and yeast extracts, 100X1mlP1021Protease inhibitor cocktail for fungal and yeast extracts, 100X5mlP1025Protease inhibitor cocktail for bacterial cell extracts, 100X1mlP1026Protease inhibitor cocktail for bacterial cell extracts, 100X5mlP1030Protease inhibitor cocktail for purification of His-tagged proteins, 100X1mlP1031Protease inhibitor cocktail for purification of His-tagged proteins, 100X5mlP1045Protease and phosphatase inhibitor cocktail for general use, 50X2ml eachP1046Protease and phosphatase inhibitor cocktail for general use, 50X10ml eachP1048Protease and phosphatase inhibitor cocktail for general use, MS-safe, 50X1ml eachP1049Protease and phosphatase inhibitor cocktail for general use, MS-safe, 50X5ml eachP1050Protease and phosphatase inhibitor cocktail for mammalian cell and tissue extracts, 50X2ml eachP1051Protease and phosphatase inhibitor cocktail for mammalian cell and tissue extracts, 50X10ml eachP1055Protease and phosphatase inhibitor cocktail for plant cell and tissue extracts, 50X2ml eachP1056Protease and phosphatase inhibitor cocktail for plant cell and tissue extracts, 50X10ml eachP1060Protease and phosphatase inhibitor cocktail for fungal and yeast extracts, 50X2ml eachP1061Protease and phosphatase inhibitor cocktail for fungal and yeast extracts, 50X10ml eachP1065Protease and phosphatase inhibitor cocktail for bacterial cell extracts, 50X2ml eachP1066Protease and phosphatase inhibitor cocktail for bacterial cell extracts, 50X10ml eachP1081Phosphatase inhibitor cocktail A, 50X2mlP1082Phosphatase inhibitor cocktail A, 50X10mlP1086Phosphatase inhibitor cocktail B ,50X2mlP1087Phosphatase inhibitor cocktail B ,50X10mlP1091Phosphatase inhibitor cocktail C ,50X2mlP1092Phosphatase inhibitor cocktail C ,50X10mlP1096Phosphatase inhibitor cocktail D ,50X2mlP1097Phosphatase inhibitor cocktail D ,50X10mlP1112Deacetylase Inhibitor Cocktail,100X1mlP1113Deacetylase Inhibitor Cocktail,100X5mlTable 2. Information of commonly used protease, phosphatase, and deacetylase inhibitors. NameMW Target EnzymeInhibition AEBSF239.5Serine proteaseirreversibleAprotinin6511.5Serine proteaseReversibleBestatin308.38AminopeptidaseReversibleE64357.4Cysteine proteaseirreversibleLeupeptin475.6Serine and Cysteine proteaseReversiblePepstatin A685.9Aspartic proteaseReversibleEDTA372.24MetalloproteinaseReversiblePhenanthroline198.22MetalloproteinaseReversiblePhosphoramidon587.5MetalloproteinaseReversibleSodium fluoride41.99Acid phosphataseReversibleSodium pyrophosphate221.9Serine/Threonine Phosphataseirreversibleβ-glycerophosphate306.11Serine/Threonine PhosphataseReversibleImidazole68.2Alkaline phosphataseReversibleSodium molybdate205.9Acid phosphataseirreversibleSodium tartrate 230.1Acid phosphataseReversibleSodium orthovanadate183.91Alkaline and Tyrosine PhosphatasesReversible(-)-p-Bromotetramisole373.22Alkaline phosphataseirreversibleCantharidin196.20Protein Phosphatase 2AReversibleMicrocystin-LR995.17Protein Phosphatase 1 and Protein Phosphatase 2AReversibleCalyculin A1009.17Protein Phosphatase 1 and Protein Phosphatase 2AReversibleTrichostatin A302.37class I/II HDACReversibleEX-527248.71SIRT1 over SIRT2 & SIRT3ReversibleNicotinamide122.12class III HDACReversibleSodium butyrate110.09class I/II HDACReversible
Instructions for Use： Protease and phosphatase inhibitors provided in this product are 50X concentrated stock solutions, and should be added to lysis buffer at a ratio of 1:50 for use (e.g., add 20μl of protease inhibitor cocktail and 20μl of phosphatase inhibitor cocktail to 1ml of lysis buffer). The lysis buffer containing protease and phosphatase inhibitors should be prepared freshly before use. If required, the 0.25M EDTA can also be added to the lysis buffer at a ratio of 1:50. After preparation, protein extraction from cells or tissues should be performed immediately.
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