| Description | Clostripain (Endoproteinase-Arg-C) is a two chain proteinase associated with collagenase and isolated from Clostridium histolyticum. It is highly specific for the carboxyl peptide bond of arginine. Clostripain has a sulfhydryl requirement; it is activated by dithiothreitol, cysteine, or other Clostripain (Endoproteinase-Arg-C) is a two chain proteinase associated with collagenase and isolated from Clostridium histolyticum. It is highly specific for the carboxyl peptide bond of arginine. Clostripain has a sulfhydryl requirement; it is activated by dithiothreitol, cysteine, or other sulfhydryl containing reagents. The presence of calcium ions is essential. The enzyme is inhibited by oxidizing agents and sulfhydryl reactants and by Co²⁺, Cu²⁺, Cd²⁺, and heavy metal ions. Citrate, borate, and Tris anions are less inhibitory.Clostripain is a cysteine-activated protease found, along with collagenase and other proteases, in culture filtrates of Clostridium histolyticum. It is unique in its specificity for the carboxyl peptide bond of arginine and its dependence on thiol and calcium ions.SpecificityClostripain selectively hydrolyzes arginyl bonds and lysyl bonds at a lower rate. It can also act as a transpeptidase with maximal activity at pH 7.6-9.0.CompositionClostripain is a heterodimer. The mature chain is composed of 526 residues. The two chains are held together by strong noncovalent forces. The catalytic sulfhydryl residue of the active site is believed to be Cys41 (heavy chain residue). The precursor contains a 27 amino acid putative signal peptide, a 23 amino acid propeptide, a 131 amino acid light chain subunit, a 9 amino acid linker peptide, and a 336 amino acid heavy chain subunit.Molecular CharacteristicsBoth the heavy and light chains are encoded by a single gene with a 1581 nucleotide open reading frame (ORF). Upon expression of the gene, the entire ORF (the signal region, proregion, and 9 amino acid peptide linker) is transcribed. Postranslational processing produces the heterodimeric active enzyme.ApplicationsPeptide mappingSequence analysisCell isolationHydrolysis/condensation of amide bondsPeptide synthesisCharacteristicsMolecular Weight: 53.0 kDa (Theoretical); Light chain: 12.5 kDa, Heavy chain: 45 kDaOptimal pH: 7.4-7.8 (activity against a-benzoyl-arginine ethyl ester)Isoelectric point: 4.8-4.9Extinction Coefficient: 87,890 cm⁻¹ M⁻¹ (Theoretical); E1%,280= 16.57 (Theoretical)Active Site Residues: Cysteine (C41, heavy chain)Activators: Sulfhydryl requirement: dithiothreitol, cysteine, or other reducing agents, Calcium ion is essential, Reducing agentsInhibitors: EDTA, Oxidizing agents, Sulfhydryl reagents (such as TLCK), Co2+, Cu2+, Cd2+, and heavy metal ions, Citrate, borate and Tris anions partially inhibitAssayMethodThe reaction velocity is measured as an increase in absorbance at 253 nm resulting from the hydrolysis of N-benzoyl-L-arginine ethyl ester. One unit hydrolyzes one micromole of BAEE per minute at 25°C and pH 7.6 under the conditions specified.Reagents0.075 M Sodium phosphate buffer, pH 7.67.5 mM Dithiothreitol (DTT)0.75 mM N-Benzoyl-L-arginine ethyl ester (BAEE)1.0 mM Calcium acetate containing 2.5 mM dithiothreitol (activation solution)EnzymeDissolve or dilute the enzyme at a concentration of 1 mg/ml in water. Immediately prior to assay, dilute the enzyme further in 1.0 mM Calcium acetate containing 2.5 mM dithiothreitol to a concentration of 0.2-0.8 units/ml.ProcedureAdjust spectrophotometer to 253 nm and 25°C.Pipette into each cuvette as follows:0.075 M phosphate buffer, pH 7.61.0 ml7.5 mM DTT1.0 ml0.75 mM BAEE1.0 mlIncubate in spectrophotometer for 3-5 minutes to achieve temperature equilibrium and establish blank rate, if any. At zero time, add 0.1 ml of appropriately diluted enzyme and record A253 for 4-5 minutes. Determine ΔA253/minute from the linear portion of the curve. Note: The reaction appears to be most linear with respect to enzyme concentration when ΔA253/min is between 0.007 and 0.030.Calculationwhere 1150 is the extinction coefficient of BAEE at 253 nm... Read More | Product IntroductionHuman epidermal growth factor (EGF) has a molecular weight of 6 KD, contains 53 amino acids, and has three intramolecular disulfide bonds.Upon binding to EGFR, a specific receptor located on the cell surface, EGF causes a series of biochemical reactions in the cell: increasing Product IntroductionHuman epidermal growth factor (EGF) has a molecular weight of 6 KD, contains 53 amino acids, and has three intramolecular disulfide bonds.Upon binding to EGFR, a specific receptor located on the cell surface, EGF causes a series of biochemical reactions in the cell: increasing the level of calcium ion concentration in the cell, promoting the progress of glycolysis, increasing the synthesis of proteins, and can enhance the expression of EGFR, a specific class of genes to promote DNA synthesis and cell proliferation.Specification parametersSource Pichia pastorisAppearance white lyophilized powderActivity ≥1.0×106IU/mgpH 6.5-7.5Molecular weight 6.5kDEndotoxin ≦1.0 EU/mgCAS No 62253-63-8Matters needing attentionReconstitution: reconstitution of REGF lyophilized powder to 100-200 µg/ml with sterile water is recommended and further dilution with other solvents may be performed.REGF dissolved at 4 ° C can be stored for 2-7 days and used up as soon as possible.To not use for short periods, store at - 20 ℃.Use as soon as possible after opening to avoid contamination.Limitations of useIt is suitable for research, laboratory and production use only and cannot be used directly in humans... Read More | FunctionSignal-transducing molecule. The receptor systems for IL6, LIF, OSM, CNTF, IL11, CTF1 and BSF3 can utilize gp130 for initiating signal transmission. Binds to IL6/IL6R (alpha chain) complex, resulting in the formation of high-affinity IL6 binding sites, and transduces the signal. Does not FunctionSignal-transducing molecule. The receptor systems for IL6, LIF, OSM, CNTF, IL11, CTF1 and BSF3 can utilize gp130 for initiating signal transmission. Binds to IL6/IL6R (alpha chain) complex, resulting in the formation of high-affinity IL6 binding sites, and transduces the signal. Does not bind IL6. May have a role in embryonic development (By similarity). The type I OSM receptor is capable of transducing OSM-specific signaling events.Post-translationalPhosphorylation of Ser-782 down-regulates cell surface expression. Heavily N-glycosylated... Read More | Purity>95% (SDS-PAGE) Endotoxin level<1.0 EU/µgFunctionInhibits the synthesis of a number of cytokines, including IFN-gamma, IL-2, IL-3, TNF and GM-CSF produced by activated macrophages and by helper T-cells | Purity:>85%, by SDS-PAGE visualized with Coomassie® Blue Staining.Description：mCherry is a bright red monomeric fluorescent protein created by rounds of directed evolution of DsRed. mCherry matures rapidly, making it possible to see results very soon after transfection or activation Purity:>85%, by SDS-PAGE visualized with Coomassie® Blue Staining.Description：mCherry is a bright red monomeric fluorescent protein created by rounds of directed evolution of DsRed. mCherry matures rapidly, making it possible to see results very soon after transfection or activation of transcription. It is highly photostable and resistant to photobleaching (Shaner et al. 2004). As a result, mCherry is now the most widely used and cited red fluorescent protein. mCherry is bright although tdTomato is the brightest commercially available red fluorescent protein... Read More |