| Description | Hydrolyzed creatinine, used in the development and bulk formulation of enzymatic creatinine reagents.Enzymatic propertiesSource: MicroorganismEnzyme Committee No. : EC 3.5.2.10Molecular weight: 29 kDa (SDS-PAGE)Isoelectric point: 5.3Km value: 5.0× 10-2 M (Creatinine),8.0× 10-2 M (Creatine)Hydrolyzed creatinine, used in the development and bulk formulation of enzymatic creatinine reagents.Enzymatic propertiesSource: MicroorganismEnzyme Committee No. : EC 3.5.2.10Molecular weight: 29 kDa (SDS-PAGE)Isoelectric point: 5.3Km value: 5.0× 10-2 M (Creatinine),8.0× 10-2 M (Creatine)Inhibitors: Hg2+, Cu2+, Fe3+Optimal pH: 7.0-8.0 Figure 1 Optimum temperature: 65℃ Figure 2pH stability: pH 5.5-10.0 (25℃, 16 h) Figure 3Thermal stability: stable below 65℃ (pH 8.0, 30 min) Figure 4Stability: -25 ~ -15℃ standing storageMaintain over 90% activity for 12 months Figure 5 Assay method for activity1. Principle 2. Definition of enzyme activityUnit enzyme activity is defined as the amount of enzyme required to catalyze a reaction to produce 1µmol creatine per minute under the following conditions.3. Reagent preparationReagent I: 0.3M potassium phosphate buffer, pH 6.5.Reagent II: 0.1M creatinine solution (1.13g creatinine dissolved in 100mL UP water).Reagent III: 4% Na2CO3 solution (4.0g Na2CO3 dissolved in 100mL UP water).Reagent IV: 2% alpha-naphthol solution (2.0g alpha-naphthol dissolved in 100 mL of 99.5% ethanol).Reagent V: 1.2g NaOH and 3.2g Na2CO3 were dissolved in double steaming water at a constant volume of 100 mL.Reagent VI: 0.05% diacetyl solution (0.05mL diacetyl with water to 100 mL).Enzyme diluent: 5 mM Tris-HCl pH 8.04. Operation procedure4.1. Add 0.1 mL reagent I and 0.8mL reagent II into a 5 mL centrifuge tube.4.2. Water bath at 37℃ for 5 minutes.4.3. Add 0.1 mL of the sample to be tested and incubate at 37℃ for 10 minutes.4.4. Add 2.0mL reagent III to terminate the reaction, remove and place on ice.4.5. Add the following reagents in the new 5 mL centrifuge tube in order:The solution for step 4: 80 µLDouble steaming water: 720 µLReagent IV: 400 µLReagent V: 400 µLReagent VI: 400 µL4.6. After standing at 25℃ for 1 h, add 2 mL of double steaming water to dilute.4.7. Use a spectrophotometer to measure the absorbance at 525 nm.* Replace enzyme liquid with enzyme diluent, other steps are the same, the absorbance of the resulting solution is blank absorbance (∆Ab)∆A=∆As- ∆Ab5. Vitality computing Vt: Total volume of reaction liquid (1.0mL);Vs: Enzyme liquid volume (0.1mL);t: Reaction time (10 minutes);df: Dilution ratio;C: Enzyme concentration (mg/mL);1.0: Optical path length (cm);0.0704: Millimolar absorption coefficient (cm²/µmol) of chromophore at 525nm under standard reaction conditions... Read More | 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). It may be used in biotechnology to cleave affinity tags from recombinant proteins, either co-translationally orin vitrofollowing purification.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). It may be used in biotechnology to cleave affinity tags from recombinant proteins, either co-translationally orin vitrofollowing purification. Its high specificity and activity at a wide range of pH and ionic strength make TEV Protease more versatile than many other proteases used for the same purpose. Unlike factor Xa, enteropeptidase or thrombin, TEV Protease has not been found to cleave at unintended sites, even when present at a high concentration. TEV Protease is a 3C-type protease that cleaves substrates with a consensus sequence of ENLYFQG. Cleavage occurs between Q and G. Since the final aa remains on the cleaved protein where it could potentially affect structure or function, substitution of a variety of aa have been tested. In order of efficiency, S, A, M, Y, D, N, E, K or L may be effectively used in place of G. Several of the remaining aa may also vary, giving a final consensus sequence of ExxYF(M)Q(E)/G(S, A or others) where aa in parenthesis are alternatives and x is any aa. The autocatalytic site of NIa at S2256 has been mutated to an N for improved stability of the protease.Tobacco Etch Virus Protease is a highly site-specific cysteine protease that is found in the tags from fusion proteins. The optimal temperature for cleavage is 30°C. It is recommended that the cleavage for each fusion protein be optimized by varying the amount of recombinant viral TEV protease, reaction time, or incubation temperature. It can be removed by Ni2+ affinity resin... Read More | Purity>95% SDS-PAGE.FunctionThe soluble form is chemotactic for T-cells and monocytes, but not for neutrophils. The membrane-bound form promotes adhesion of those leukocytes to endothelial cells. May play a role in regulating leukocyte adhesion and migration processes at the endothelium. Binds toPurity>95% SDS-PAGE.FunctionThe soluble form is chemotactic for T-cells and monocytes, but not for neutrophils. The membrane-bound form promotes adhesion of those leukocytes to endothelial cells. May play a role in regulating leukocyte adhesion and migration processes at the endothelium. Binds to CX3CR1.Post-translationalA soluble short 95 kDa form may be released by proteolytic cleavage from the long membrane-anchored form. O-glycosylated with core 1 or possibly core 8 glycans... Read More | Fibronectin (FN) is a particularly important and well-studied component of the extracellular matrix, and is known to play a key role in cell adhesion, growth, spreading, migration, differentiation and proliferation. Fn is a 200-250 kDa glycoprotein composed of 2 subunits bound via a disulfide bond. Fibronectin (FN) is a particularly important and well-studied component of the extracellular matrix, and is known to play a key role in cell adhesion, growth, spreading, migration, differentiation and proliferation. Fn is a 200-250 kDa glycoprotein composed of 2 subunits bound via a disulfide bond. Currently, the Fn is purified from the plasma, which however is limited by the availability of supply. The the recombinant human fibronectin (OsrhFN) was expressed in the rice endosperm platform, which is animal component free and has high purity, and has been demonstrated has the same physical and chemical with the plasma derived Fn. OsrhFN provides a safety solution to replace the plasma derived FN.pH value: 6.0-8.0... Read More | Purity>95% by SDS-PAGE and HPLC analyses.FunctionLigand for IL17RA and IL17RC (PubMed:17911633). The heterodimer formed by IL17A and IL17F is a ligand for the heterodimeric complex formed by IL17RA and IL17RC (PubMed:18684971). Involved in stimulating the production of other cytokines such as IL6Purity>95% by SDS-PAGE and HPLC analyses.FunctionLigand for IL17RA and IL17RC (PubMed:17911633). The heterodimer formed by IL17A and IL17F is a ligand for the heterodimeric complex formed by IL17RA and IL17RC (PubMed:18684971). Involved in stimulating the production of other cytokines such as IL6, IL8 and CSF2, and in regulation of cartilage matrix turnover (PubMed:11591732, PubMed:11591768, PubMed:11574464). Also involved in stimulating the proliferation of peripheral blood mononuclear cells and T-cells and in inhibition of angiogenesis (PubMed:11591732). Plays a role in the induction of neutrophilia in the lungs and in the exacerbation of antigen-induced pulmonary allergic inflammation... Read More |