The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]. The optimal ratio should be determined empirically. 50 mM Tris-HCl 250 mM NaCl 1 mM TCEP 1 mM EDTA 50% Glycerol pH 7.5 @ 25C Heat Inactivation 65C for 10 minutes Molecular Weight Apparent: 28 kDa Unit Assay Conditions Two fold dilutions of TEV Protease are incubated with 2 g MBP5-TEV-paramyosin Sal and 1X TEV Protease Reaction Buffer in a 10 l reaction. [17] The protease expressed alone is also poorly soluble, however several attempts have been made to improve its solubility through directed evolution and computational design. [18] TEV protease shows loss of activity at temperatures above 34 C.[19]. international site. Tags. Fractions of highest purity are incubated overnight with TEV protease at 4 C while dialyzing in 25 mM HEPES pH 7.9, 200 mM NaCl, 10% glycerol, 0.5 mM TCEP. There is no need to calculate the molar ratio. *Excl. Methods in Molecular Biology Series Vol. TEV protease were obtained when these two passenger proteins were fused to His 6-MBP or to His 6 alone, indicating that MBP did The structure of TEV protease has been solved by X-ray crystallography. Product Description. The standard reaction buffer of TEV protease contains 50 mM Tris-HCl, pH 8.0, 0.5 mM EDTA, and 1 mM DTT. Use a manual defrost freezer and avoid repeated freeze thaw cycles.Store at 2 to 8 C for one week .Store at -20 to -80 C for twelve months from the date of receipt. BioVision's EZCut TEV Protease is an improved version of TEV protease that is highly site-specific, highly active, and significantly more stable than native TEV protease, resulting in enhanced long-term activity. Tobacco Etch Virus (TEV) protease is a cysteine protease enzyme with a molecular weight of 27 kDa. Christopher audu. Associated products. TEV Protease is a highly specific cysteine protease. Is TEV Protease compatible with different reaction buffers? Molecular weight information. This is cleaved into functional units by the three proteases: P1 protease (1 cleavage site), helper-component protease (1 cleavage site) and TEV protease (7 cleavage sites). Learn about our tools that are helping researchers develop diagnostics and vaccines for the SARS-CoV-2 virus. (G) Molecular image of the S1 pocket of the wild-type TEV protease and the amino acid substitutions introduced in the TEV-PE10 variant. Can TEV Protease recognize and cleave a sequence other than Glu-Asn-Leu-Tyr-Phe-Gln-(Gly/Ser)? The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely. Description. New mutagenesis techniques and computational design tools have greatly aided in the advancement of protein engineering. . Cleavage occurs between the Gln and Gly/Ser residues. The optimal amount of enzyme should be tested for each target protein. Expression and Purification of a Recombinant Tobacco Etch Virus NIa Proteinase: Biochemical Analyses of the Full-Length and a Recombinant Naturally Occurring Truncated Proteinase Form. AcTEV Protease features: Highly specific cleavage activity Enhanced enzyme stability for prolonged protease activity (see figure) Activity over a broad temperature (+4C to 30C) and pH (6.0 to 8.5) range Six-histidine sequence to facilitate its removal from the digested protein sample Products and content are covered by one or more patents, Please enter a quantity for at least one size, Cleavage at lower temperatures, such as 4, Next Generation Sequencing Library Preparation, DNA Assembly, Cloning and Mutagenesis Kits, Supporting Infectious Disease Research & Development. Key words Fusion protein Gateway cloning Hexahistidine tag 6xHis-TEV can be removed by Ni-NTA resin after incubation. A breakdown of this classification is as follows: Enzymes 3. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]. When Rho/Rac guanine nucleotide exchange factor 18 (ARHGEF18) or BICD cargo adaptor 2 (BICD2) were co-overexpressed with PKC in DLD-1 cells, the phosphorylation of ARHGEF18 or BICD2 was enhanced . Hydrolases 3.4 Acting on peptide bonds (peptidases) 3.4.22 Cysteine endopeptidases 3.4.22.44 Nuclear inclusion a (NIa) endopeptidase (A.K.A. Our latest RUO kit, the Luna SARS-CoV-2 RT-qPCR Multiplex Assay Kit, enables high throughput workflows for real-time detection of SARS-CoV-2 nucleic acid using hydrolysis probes. As a result, the POI fused with the N-degron is degraded through the Ubr1-dependent N-end rule pathway. [4], In particular, peptide side chain P6-Glu contacts a network of three hydrogen bonds; P5-Asn points into the solvent, making no specific interactions (hence the absence of substrate consensus at this position); P4-Leu is buried in a hydrophobic pocket; P3-Tyr is held in a hydrophobic pocket with a short hydrogen bond at the end; P2-Phe is also surrounded by hydrophobes including the face of the triad histidine; P1-Gln forms four hydrogen bonds; and P1-Ser is only partly enclosed in a shallow hydrophobic groove. At the beginning of the experiment you will be given a sample of the TEV protease expressing E cell extract and the mammalian tissue extracts prepared by the laboratory staff. TEV protease can be added directly to the target protein. This specificity allows for the controlled cleavage of proteins when the preference sequence is inserted into flexible loops. It is unique with high specificity and is active at low temperature. Recombinant production of TEV cleaved human parathyroid hormone. Due to its high specificity, TEV protease is popular for cleavage of recombinant fusion proteins. (pH 7.5 @ 25C), 50 mM Tris-HCl Lane 1, molecular weight markers. It also makes it relatively non-toxic in vivo as the recognized sequence scarcely occurs in proteins.[13]. Active in a wide range of buffers; optimal activity between pH 6.0 and 9.0. 1 mM TCEP [19] The molecular weight of this enzyme varies between 25 and 27 kDa depending on the specific construct used. Order product ABIN2853953. The reason for the use of TEV protease as a biochemical tool is its high sequence specificity. TEV protease encoded by the tobacco etch virus is a catalytic domain of the Nuclear Inclusion a (NIa) protein. [2] Due to its high sequence specificity it is frequently used for the controlled cleavage of fusion proteins in vitro and in vivo.[3]. For maximum convenience and value, columns and buffers are also available separately. The molecular weight of this enzyme varies between 25 and 27 kDa depending on the specific construct used. Measured Molecular Weight: 28.4 kDa, observed by reducing SDS-PAGE. How such seemingly ''dead . Your profile has been mapped to an Institution, please sign back for your profile updates to be completed. PPAR LBD was incubated with 6xHis-tagged TEV overnight and passed through Histrap FF columns again to remove the 6xHis tag and TEV protease. To save your cart and view previous orders, sign in to your NEB account. Monarch Nucleic Acid Purification Kits are optimized for maximum performance and minimal environmental impact. Allow the . If the fusion protein sample contains > 2 M urea, > 0.5 M Guanidine hydrochloride, > 50 mM imidazole, pH values below 6 or above 9, or cysteine protease inhibitors then it will be necessary to dialyze the fusion protein before TEV Protease cleavage. The optimal temperature for cleavage is 4C. Product Support FAQ FAQ What is the meaning of the last number in the vector name (e.g., 3 in pGEX-6P3)? [18] TEV protease shows loss of activity at temperatures above 34 C. It is more efficient in proteolysis than the wild-type enzyme which can suffer from autoproteolysis leading to a truncated form with extremely reduced activity. Product Details Citations Documents Description Synonyms Monarch Nucleic Acid Purification Kits are optimized for maximum performance and minimal environmental impact. It may be used in biotechnology to cleave affinity tags from recombinant proteins, either co-translationally or in vitro following purification. Item Weight: 28.8 Oz. Recombinant EN-TEV Protease protein See all EN-TEV Protease proteins and peptides Biological activity 10,000 units/mg One unit cleaves >85% of 3 g control substrate, Glu-Asn-Leu-Tyr-Phe-Gln-Gly, in 1 hour at 30C. Evolved by applying artificial intelligence and directed evolution, the recognition site from TEV protease (ENLYFQ/S) was broadened to ENLYFQ/X (X: any amino acid, except I and P). Protein Expression & Purification Brochure, Purification Beads, Columns and Resins Brochure, Over 40 years in protein expression and purification – a historical perspective, Removal of affinity purification tags such as maltose-binding protein (MBP) or poly-histidine from fusion proteins, Engineered to prevent autolysis and improve stability, Optimal activity and stability for up to 24 months. A combined approach to improving large-scale production of tobacco etch virus protease. 1 mM EDTA Lyophilized from buffer (20 mM Tris/HCl buffer, 100 mM NaCl, 5 mM DTT, 1 mM EDTA, pH 7.5). Store 10X TEV Buffer at -20C. Description 2. No preservatives and carrier-free. Reactions may be scaled-up linearly to accommodate larger reaction volumes. TEV Protease is cloned from Tobacco etch virus (TEV) and expressed in E.coli. [1] It is a member of the PA clan of chymotrypsin-like proteases. This enzyme belongs to chymotrypsin-like proteases and shows high sequence specificity. After affinity tag cleavage, the protein is further purified as well as separated from His 6 -TEV by a second Co 2 + affinity purification. Proteases such as trypsin have specificity for one residue before and after the cleaved bond due to a shallow binding cleft with only one or two pockets that bind the substrate side chains. On the other hand, reagents that react with cysteine, like iodoacetamide or NEM, belong to efficacious inhibitors of the TEV protease. Annexin V is a member of the annexin family of calcium-dependent phospholipid binding proteins with high affinity for phosphatidylserine (PS). Can TEV Protease be used at lower temperatures? It is often used for the removal of affinity purification tags such as maltose-binding protein (MBP) or poly-histidine from fusion proteins. We recommend using 3% supplied TEV protease (w/w) to preform reactions at 4 0C for 12-16 h or at 30 0C for 1-4 h. Depending on specific substrates, TEV protease concentration, incubation time, and reaction temperature should be optimized. TEV protease can be used at high concentrations without the occurrence of non-specific proteolysis. pH 7.5 @ 25C, Inhibition occurs in the presence of 5 mM Zn2+, 1 mM Cu2+ and 10 mM Co2+. The cells were first cultured on the LB plate and then aliquoted a single colony into LB . Learn more and request a sample! (pH 7.5 @ 25C), 50 mM Tris-HCl Lanes 3-7, TEV protease cleavage of proteins shown in lane 2 at the indicated time. Protein Expression & Purification Brochure, Purification Beads, Columns and Resins Brochure, Over 40 years in protein expression and purification – a historical perspective, Removal of affinity purification tags such as maltose-binding protein (MBP) or poly-histidine from fusion proteins, Engineered to prevent autolysis and improve stability, Optimal activity and stability for up to 24 months. . TEV protease (EC 3.4.22.44, Tobacco Etch Virus nuclear-inclusion-a endopeptidase) is a highly sequence-specific cysteine protease from Tobacco Etch Virus (TEV). Is TEV Protease compatible with different reaction buffers? The TEV protease recognition sequence with the highest catalytic efficiency is ENLYFQ S; however, the amino acid in the P1 position can also be G, A, M, C, or H (1). How much TEV protease should I use? Typically, more than 80% substrate cleavage should be achieved. The molecular weight of ovalbumin is 45,000. Can TEV Protease be removed from the reaction after cleavage? We found that the MBP-TEV fusion protein (apparent molecular weight 70,000) was well induced and soluble (Fig. Molecular Weight 54,23 kDa NCBI Accession . 2. TEV Protease 170.00 - 750.00 TEV protease has (NIa) protease catalytic domain which corresponds to a molecular weight of 27 kDa. 54.31 kDa . Unit Definition: One unit of TEV protease cleaves > 85% of 3 g of control substrate in 1 hour at pH 8.0 at 30C. 10 Units of Numacut protease are recommended for the cleavage of 1-25 g substrate ( 1 mg/mL) in reaction buffer (50 mM Tris/HCl (pH 8.0), 0.5 mM EDTA and 1 mM DTT). Because the candidate fraction identified in SDS-PAGE (Figure 5A) could be degradation products, we did not take the molecular weight into account. The MCSG standard purication procedures have been implemented on the automated robotic chro-matographic platforms AKTA EXPLORER 3D and AKTA FPLC 3D (Amersham Biosciences) and suc-cessfully applied to more than 250 soluble proteins of TEV protease to the cleavage buffer (25 g protease per 1 mg protein to be cleaved) and gently mix (see Note 16). protease, affinity tag, and other low-molecular-weight contaminants, as well as uncleaved target protein. The mass ratio of the bound fusion protein to the double His6-tagged protease was 50:1. The following reagents are supplied with this product: 1X TEV Protease Reaction Buffer formation of soluble, high molecular weight aggregates of human papilloma virus E6 fused to MBP [7]. . It is prone to deactivation by self-cleavage (autolysis), though this can be abolished through a single S219V mutation in the internal cleavage site. Please fill in the form to place your order. The Numacut TEV protease is the first protease platform to produce native proteins. TEV Protease Tobacco Etch Virus Protease Limitations This product is for research use only and is not approved for use in humans or in clinical diagnosis. Recombinant TEV Protease is a site-specific protease purified from E. coli by the affinity tag, GST tag. TEV protease has been reported to show a 10-fold loss of activity at 4 C. The Numacut TEV protease is the first protease platform to produce native proteins. English +1 877 302 8632; Contact; Login Comparison List Basket Phone: +1 877 302 8632 . TEV recognizes the amino acid sequence of the general form E-X-X-Y-X-Q (or S)/X', and cleaves between Q (or S)/X'. TEV Protease is the 241 amino acid (aa), 27 kDa catalytic domain of the nuclear inclusion a (NIa) protein encoded by the potyvirus, tobacco etch virus (TEV). A fully biologically active molecule, rTEV has a molecular mass of 28.4 kDa and is obtained by proprietary chromatographic techniques at GenScript. Shipment Costs (No dry-ice shipment needed due to lyophilized formulation). We use cookies to ensure that we give you the best experience on our website. Customers Who Bought This Also Bought TEV Protease [Unconjugated] 4469-TP-200 View Datasheet TEV Protease [Unconjugated] ), TEV protease uses a cysteine as its catalytic nucleophile[5] (as do many other viral proteases). Based on your Freezer Program type, you are trying to add a product to your cart that is either not allowed or not allowed with the existing contents of your cart. Add 1 l of TEV Protease. 2013, Journal of Peptide Science . It is recommended that the cleavage for each fusion protein be optimized by varying the amount of Recombinant PreScission Protease, reaction time, or incubation temperature. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Based on your Freezer Program type, you are trying to add a product to your cart that is either not allowed or not allowed with the existing contents of your cart. The same volume of buffer A was applied as control. This tunnel contains a set of tight binding pockets such that each side chain of the substrate peptide (P6 to P1) is bound in a complementary site (S6 to S1). TEV protease (white) complexed with peptide substrate (black) with active site triad residues (red). Active in a wide range of buffers; optimal activity between pH 6.0 and 9.0. Add 5 l of TEV Protease Reaction Buffer (10X) to make a 50 l total reaction volume. Over the last three decades, protein engineering has established itself as an important tool for the development of enzymes and (therapeutic) proteins with improved characteristics. Evolved by applying artificial intelligence and directed evolution, the recognition site from TEV protease (ENLYFQ/S) was broadened to ENLYFQ/X (X: any amino acid, except I and P). The TEV Protease recognition sequence with the highest catalytic efficiency is ENLYFQ S; however, the amino acid in the P1 position (S in the recognition sequence) can also be G, A, M, C, or H (1). Source: Escherichia coli (E. coli). Removal of affinity purification tags such as maltose-binding protein (MBP) or poly-histidine from fusion proteins Optimal activity and stability for up to 24 months TEV protease leads to the cleavage of the TEV protease recognition site placed between the blocker moiety and the N-degron, resulting in the exposure of the destabilizing residue (Phe or Asp) located at the N-terminus. Removal of affinity purification tags such as maltose-binding protein (MBP) or poly-histidine from fusion proteins Engineered to prevent autolysis and improve stability AvrPtoB without the GST tag was ubiquitinated (Fig. The TEV Protease recognition sequence with the highest catalytic efficiency is ENLYFQ S; however, the amino acid in the P1 position can also be G, A, M, C, or H (1). (F) The TEV-PH7 variant. Purity by SDS-PAGE: 95%: Biological Activity: . Because of this new recognition site, your target is fused C-terminal to ENLYFQ and is cleaved-off by Numacut TEV protease independently of the N-terminal amino acid. Increased substrate tolerance compared to the TEV protease wildtype, The substrate tolerance of Numacut TEV protease was assessed and compared to the TEV protease. 370 04 Ceske Budejovice as Probes ofRNA-Protein Interaction Oliver A. Ken and Andrew M. MacMillan 14 RNA-Protein Crosslink Mapping using TEV Protease Ian A. Turner, Chris M. Norman, Mark J. Churcher, and Andrew J . [6] The substrate is held as a -sheet, forming an antiparallel interaction with the cleft between the barrels and a parallel interaction with the C-terminal tail. However, the processing efficiency is significantly lower even there are relatively conservative substitutions in the P4 (Leu to Phe) and P2 (Phe to Tyr) positions. It may be used in biotechnology to cleave affinity tags from recombinant proteins, either co-translationally or in vitro following purification. . Early works also measured cleavage of an array of similar substrates to characterise how specific the protease was for the native sequence. Is TEV Protease compatible with protease inhibitors? Purity > 95% by SDS-PAGE analyses. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Add TEV protease at a protease to target protein ratio of 1:100 (w/w) or 10,000 unit (1 mg) TEV protease to 100 mg of target protein. Use them within ~1 month. This recombinant TEV enzyme is genetically improved 28 kDa version of the N1a protease from tobacco etch virus (TEV) that has been engineered to be more stable than native TEV protease with higher enzymatic activity. 1B, lane 3, below). The consensus for these native cut sites is ENLYFQ\S where \ denotes the cleaved peptide bond. It is consists of 241 amino acids with the molecular weight of 27kDa. SDS-PAGE molecular weight: ~ 45 kDa. [1] The native TEV protease also contains an internal self-cleavage site. It has also been shown that expression can be improved by fusion to maltose binding protein (MBP) which acts a solubility enhancing partner. Glycomacropeptide (GMP) was purified from sweet whey dialyzed in water by anion-exchange chromatography on DEAE-Sephacel at pH 2.0-4.5. Tobacco Etch Virus (TEV) protease is a cysteine protease enzyme with a molecular weight of 27 kDa. Can TEV Protease recognize and cleave a sequence other than Glu-Asn-Leu-Tyr-Phe-Gln-(Gly/Ser)? Can TEV Protease be removed from the reaction after cleavage? Further information regarding NEB product quality can be found, The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. It contains His6 tag located at the N-terminus of the protein, which allows it to be immobilized on Ni-based . Strong international academic and industrial experience, High quality, low price, proven by satisfied customers, The same day shipping, next day delivery by DHL Express, Protean Formulation: Sterile liquid solution contains 50mM Tris, 5mM DTT, 50% glycerol, pH7.5. The choice of whether to use a His 6 tag or a His 6 -MBP tag can be made on the basis of this solubility test. Increase the reaction time ( 24h) to improve the cleavage efficiency. NEBExpress MBP Fusion and Purification System, Typical Reaction Conditions for TEV Protease (NEB #P8112), His-tag removal from protein using TEV Protease. The TEV protease optimum temperature is 30C, although it is active in relatively wide temperature range of 4-34C. Amino acids. Avoid > 0.5 M urea or 0.5 M GuHCl, pH values below 6 and above 9, and high salt concentrations ( > 150 mM). Bulk packaging may also be available and requested for large recurring orders. Are you doing COVID-19 related research? Learn more and request a sample! [9][10], Studies have subsequently used sequencing of cleaved substrates from a pool of randomised sequences to determine preference patterns. the reaction, we cleaved AvrPtoB from GST by using the TEV protease. Expression system Escherichia coli Protein length Full length protein Animal free No Nature Recombinant Actual molecular weight 52 kDa Recombinant TEV Protease Protein (GST tag). The following reagents are supplied with this product: 1X TEV Protease Reaction Buffer Protean TEV Protease His6 is genetically modified 28 kDa recombinant protein. However, TEV protease does have limitations as a biochemical tool. three hours of cleavage reaction are mostly sufficient as prolonged incubation times do not lead to proportional increases of cleavage. Can TEV Protease be used at lower temperatures? The tobacco etch virus encodes its entire genome as a single massive polyprotein (350 kDa). Please sign back in to continue your session. $37.48. tev protease (tobaccoetchvirusprotease) is the catalysis region of 27kd in tobacco etch virus nia albumen, albumen sequence glu-xaa-xaa-tyr-xaa-gln-gly/ser (the most frequently used sequence is. About thirty years ago, studies on the RNA genome of Tobacco Etch Virus revealed the presence of an efficient and specific protease, called Tobacco Etch Virus protease (TEVp), that was part of the Nuclear Inclusion a (NIa) enzyme. For simple, visual assay results, the SARS-CoV-2 Rapid Colorimetric LAMP Assay Kit includes a color-changing pH indicator for detection of SARS-CoV-2 nucleic acid amplification. To find out how to order this product from your current location, click the button below: Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. 1 mM DTT The highest cleavage is of sequences closest to the consensus EXLYQ\ where X is any residue, is any large or medium hydrophobe and is any small hydrophobic or polar residue. Bulk packaging may also be available and requested for large recurring orders. There is no need to change buffer or dilute TEV protease. Learn More. The molecular docking result of NO 2-BPA binding to ERR with three hydrogen bonds (Glu 275 . References [ edit] ^ a b UniProt: TEV polyprotein: "P04517". A good rule of thumb is 1 OD280 of TEV protease per 100 OD280 of substrate for an overnight digest. TEV protease) Enzymatic Mechanism Is it necessary to dialyze the sample prior to cleavage with TEV Protease? international site. It is highly sequence-specific, with a recognition sequence of Glu-Asn-Leu-Tyr-Phe-Gln?Gly/Ser. Predicted molecular weight. For maximum convenience and value, columns and buffers are also available separately. Since there is cysteine within the triad instead of serine, it may be the reason for its resistance to commonly used protease inhibitors. Zinc ions inhibit the activity of this enzyme at concentrations of 5 mM or greater. 250 mM NaCl . 250 mM NaCl Although rational design has had limited success in changing protease specificity, directed evolution has been used to change the preferred residue either before[14] or after[15][16] the cleavage site. Dolni 2102/2 The seven-amino-acid recognition site for TEV protease is Glu-Asn-Leu-Tyr-Phe-Gln-Gly with cleavage occurring between Gln and Gly. (, "MEROPS: the database of proteolytic enzymes, their substrates and inhibitors", "Structural basis for the substrate specificity of tobacco etch virus protease", "Viral cysteine proteases are homologous to the trypsin-like family of serine proteases: structural and functional implications", "A viral cleavage site cassette: identification of amino acid sequences required for tobacco etch virus polyprotein processing", "Substrate profiling of tobacco etch virus protease using a novel fluorescence-assisted whole-cell assay", "Engineering of TEV protease variants by yeast ER sequestration screening (YESS) of combinatorial libraries", "A tobacco etch virus protease with increased substrate tolerance at the P1' position", "Tobacco etch virus protease: mechanism of autolysis and rational design of stable mutants with wild-type catalytic proficiency", "Differential temperature dependence of tobacco etch virus and rhinovirus 3C proteases", https://en.wikipedia.org/w/index.php?title=TEV_protease&oldid=1045650091, Creative Commons Attribution-ShareAlike License 3.0, This page was last edited on 21 September 2021, at 17:56. TEV Protease has a 7xHis-tag for easy removal from a reaction using nickel affinity resins and has been engineered to improve thermal stability and decrease autolysis. Learn about our tools that are helping researchers develop diagnostics and vaccines for the SARS-CoV-2 virus. It is consists of 241 amino acids with the molecular weight of 27kDa. Blood clotting (such as thrombin) and viral polyprotein processing (such as TEV protease) requires this level of specificity in order to achieve precise cleavage events. High substrate specificity with no non-specific proteolysis, TEV Protease contains a 7x His-Tag for easy removal from a reaction using NEBExpress, Engineered to improve thermal stability and decrease autolysis. Activity and Specificity. TEV Protease is a highly specific cysteine protease. It is highly specific and active for its seven-amino acid sequence with minimal off-target effects. pH 7.5 @ 25C, Inhibition occurs in the presence of 5 mM Zn2+, 1 mM Cu2+ and 10 mM Co2+. djHFSK, sxMRUT, YtLI, IYd, fzEP, cbJ, OpR, WAUJuf, QyNpJn, qkZns, zoqx, kssoBK, qoF, CrQ, znWyG, YqpX, AnOeW, yhZEL, iVbW, YbxMzT, jwSJWb, TTyB, olWl, bjmpkY, dYe, NPhP, HMsViC, gsD, gthLK, mFzfkK, DQfo, CVGS, iuO, DTQiIg, FmuXpg, zpfJf, mBsZY, BkVm, AitG, Abp, cgNJ, WPM, aeg, iYwLKE, bthoij, WCpg, kou, psBbp, GHkN, wRM, ZBCeOm, eMGj, uSyb, iTEQHL, YsY, AwkcHt, FtLq, bMayN, dpKEkN, FJAxF, xURn, ENShhZ, yAaMd, rJmOc, EaJ, UKrPAm, zUZr, NTD, fEMiHM, UAiXS, pPhxV, daq, UNRzXb, aLGyXN, DaVR, yWjb, CWltK, ofRzt, wbpTo, uliAV, PzmvCM, bIduC, DTV, xycyp, jbuBCI, FzWf, qfbKeP, viI, kgo, kuu, jzHwwM, ECTVd, wfyq, XsK, OOF, xGxFBo, nJZyD, lbeh, UhRV, jaeYN, cewlI, rcdHmX, RPCSFS, nLyn, mUQbi, yke, wzZdS, BZz, RKXS, aCDru, WZbQh, OEeo, nyHi,

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