| Description | ApplicationFor the detection of lactic acid content.Enzymatic propertiesSource: MicroorganismEnzymology Committee Number: EC 1.1.3.2Molecular weight: 42 kDa (SDS-PAGE)Isoelectric point: pH 4.6Km value: 7.5 × 10-4 M (L-Lactate)Inhibitors: Hg²⁺, Pb²⁺ Optimum pH: 6.0-7.0 ApplicationFor the detection of lactic acid content.Enzymatic propertiesSource: MicroorganismEnzymology Committee Number: EC 1.1.3.2Molecular weight: 42 kDa (SDS-PAGE)Isoelectric point: pH 4.6Km value: 7.5 × 10-4 M (L-Lactate)Inhibitors: Hg²⁺, Pb²⁺ Optimum pH: 6.0-7.0 Figure 1 Optimum temperature: 50℃ Figure 2pH stability: 6.0-8.5 (25℃,16h) Figure 3 Thermal stability: Stable below 50℃ (pH7.0, 30min) Figure 4Stability: -25 ~ -15℃ standing store for 12 monthsMore than 90% activity Figure 5 Enzyme activity definitionUnit enzyme activity is defined as the amount of enzyme required to catalyze the production of 1µmol H2O2 per minute under the following conditions.Assay method for activity1. PrincipleThe amount of Quinoneimine dye produced by the reaction can be measured by spectrophotometer at 555 nm.2. Definition of enzyme activityUnit enzyme activity is defined as the amount of enzyme required to catalyze the production of 1 µmol H2O2 per minute under the following conditions.3. Reagent preparationReagent I: 0.2 MpH 6.5 potassium phosphate buffer.Reagent II: 1kU/mL peroxidase (POD) solution.Reagent III: 50 mM4-AA solution.Reagent IV: 0.5 MDL- lactic acid solution, pH6.5.Reagent V: 50 mMTOOS solution.Enzyme diluent: 10 mMpH7.0 potassium phosphate buffer containing 10 µM FAD.Sample: Dilute the enzyme with enzyme diluent to 0.05-0.2U/mL.Prepare the reaction mixture as follows:Reagent I is 10 mlReagent II 0.25 mLReagent III 1.5 mLReagent IV is 5 mLReagent V 1.5 mLDouble steam water to 50 ml4. Operation procedure4.1 Add 1mL reaction mixture into 1mL colorimetric dish.4.2 Preheat the reaction mixture at 37 °C for 5min.4.3 Add 20µL of enzyme liquid to be measured and mix well.4.4 The reaction is measured at 37 °C at 555 nm and the absorbance change (∆As) within 1min is recorded.* 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 computing1.020: total volume of reaction liquid (mL);0.020: enzyme liquid volume (mL);1: optical path length (cm);1/2:1 mole of hydrogen peroxide to generate 1/2 mole of quinone imide dye;df: dilution ratio;C: Enzyme concentration (mg/mL);39.2: Under standard reaction conditions, the millimolar absorption coefficient of the color group at 555 nm (cm2/µmol)... Read More | Es Taq DNA Polymerase is an optimized mixed enzyme of Taq and Pfu DNA Polymerase, with 5 '→ 3' DNA polymerase activity, 5 '→ 3' exonuclease activity, and 3 '→ 5' exonuclease activity. Compared with Taq DNA Polymerase, Es Taq DNA Polymerase has excellent performance of high Es Taq DNA Polymerase is an optimized mixed enzyme of Taq and Pfu DNA Polymerase, with 5 '→ 3' DNA polymerase activity, 5 '→ 3' exonuclease activity, and 3 '→ 5' exonuclease activity. Compared with Taq DNA Polymerase, Es Taq DNA Polymerase has excellent performance of high amplification efficiency and low mismatch rate, and can efficiently amplify DNA fragments. Most of the PCR products amplified with this product contain an "A" base at the 3 'end, which can be directly used for T/A cloning. This product is suitable for conventional PCR reactions and gene cloning reactions that require high fidelity. E665597Component500 UStorageE665597AEs Taq DNA Polymerase, 5 U/µL 100 µL -20℃. Avoid freeze/thaw cycle.E665597B10×PCR Buffer 1.8 mL -20℃. Avoid freeze/thaw cycle.Activity definition:Using activated salmon sperm DNA as a template/primer, the amount of enzyme required to incorporate 10 nmol of deoxyribonucleotide into acidic insoluble substances is defined as 1 active unit (U) at 74 ℃ for 30 minutes.Quality control:After multiple column purifications, SDS-PAGE detected a purity of over 99%; No exogenous nuclease activity detected; PCR method for detecting residual DNA without host; Can effectively amplify single copy genes in the human genome; Store at room temperature for one month without significant changes in activity.1. PCR reaction system Reagent 50 µlReaction system Final concentration 10×PCR Buffer 5 µL 1× dNTP Mix，10 mM each 1 µL 200 µM each Forward Primer，10 µM 2 µL 0.4 µM Reverse Primer，10 µM 2 µl 0.4 µM Template DNA <0.5 µg <0.5 µg/50 µl Es Taq DNA Polymerase，5 U/µl 0.25-0.5 µl 1.25-2.5U/50 µl ddH2O up to 50 µL /Attention: The primer concentration should be between 0.1 and 1.0 as the final concentration µ M serves as a reference for setting the range. In the case of low amplification efficiency, the concentration of primers can be increased; When non-specific reactions occur, the primer concentration can be reduced to optimize the reaction system. 2. PCR reaction conditions Step Temperature Time / Pre denaturation 94℃ 2 min / Denaturation 94℃ 30 s 25-35 cycles Anneal 55-65℃ 30 s 25-35 cycles Extend 72℃ 30 s 25-35 cycles Finally extended 72℃ 2 min / Attention:1) In general experiments, if the annealing temperature is 5 ℃ lower than the melting temperature Tm of the amplification primer, and the ideal amplification efficiency cannot be achieved, the annealing temperature should be appropriately reduced; When non-specific reactions occur, increase the annealing temperature to optimize the reaction conditions.2) The extension time should be set according to the size of the amplified fragment. The amplification efficiency of Es Taq DNA Polymerase in this product is 2 kb/min.3) The number of cycles can be set based on the downstream application of the amplification product. If the number of cycles is too small, the amplification amount is insufficient; If there are too many cycles, the probability of mismatches will increase, and non-specific backgrounds will be severe. So, while ensuring product yield, the number of cycles should be minimized as much as possible... Read More | Inquire | Purity:>95%, by SDS-PAGE visualized with Coomassie® Blue Staining. Description： 100B, previously called S100 beta, belongs to the S100 family within the EF-hand superfamily of Ca2+ binding proteins. S100 proteins contain two EF-hand motifs that differ in affinity, separated by a hingePurity:>95%, by SDS-PAGE visualized with Coomassie® Blue Staining. Description： 100B, previously called S100 beta, belongs to the S100 family within the EF-hand superfamily of Ca2+ binding proteins. S100 proteins contain two EF-hand motifs that differ in affinity, separated by a hinge region with a hydrophobic cleft that is exposed upon Ca2+ binding. S100B is a 91 amino acid (aa) protein, after removal of the initial methionine, and is found as homodimers of 10.4 kDa monomers. Human S100B shares 99%, 98%, 100%, 99% and 97% aa sequence identity with mouse, rat, rabbit, equine and bovine S100B, respectively. Within the S100 family, human S100B shows the highest aa identity (59%) with S100A1. S100B is expressed primarily by astrocytes and oligodendrocytes in the central nervous system, and by Schwann cells in the peripheral nervous system. Ca2+-bound S100B interacts in vitro with at least 20 cytoplasmic proteins, including several structural molecules such as tubulin and GFAP. It can inhibit the phosphorylation of these kinase substrates and others such as tau and neuromodulin. Astrocytes can secrete S100B, which then acts in a cytokine-like manner. Nanomolar concentrations of S100B are secreted constitutively, promote proliferation, and are neurotrophic and anti-apoptotic. Blood levels of S100B reflect extracellular concentrations within the nervous system, and are elevated in Down’s syndrome, Alzheimer’s disease and Tourette’s syndrome, metabolic stress, acute brain injury and brain tumors. Micromolar concentrations of S100B can be destructive and pro-apoptotic; they induce the expression of iNOS, COX-2, IL-1, IL‑6 and TNF-alpha by microglia, astrocytes or neurons. Most extracellular actions of S100B can be mediated by RAGE (receptor for advanced glycation end products), which is also a receptor for other S100 proteins... Read More | Tankyrase-IN-2 (compound 5k) is a potent, selective, and orally active tankyrase inhibitor ( IC 50 s of 10, 7, and 710 nM for TNKS1, TNKS2 as well as PARP1, respectively). Tankyrase-IN-2 has favorable physicochemical profile and pharmacokinetic properties modulating Wnt pathway activity in a Tankyrase-IN-2 (compound 5k) is a potent, selective, and orally active tankyrase inhibitor ( IC 50 s of 10, 7, and 710 nM for TNKS1, TNKS2 as well as PARP1, respectively). Tankyrase-IN-2 has favorable physicochemical profile and pharmacokinetic properties modulating Wnt pathway activity in a colorectal xenograft modelIn VitroTankyrase-IN-2 (1-10000 nM; 24 hours) leads to a dose-dependent increase of tankyrase protein abundance with an EC 50 of 320 nM in DLD1 cells. This is in the same potency range as the value for axin2 increase (EC 50 =319 nM). MCE has not independently confirmed the accuracy of these methods. They are for reference only.IC50& Target:IC50: 10 nM (TNKS1), 7 nM (TNKS2), 710 nM (PARP1)... Read More |