| Description | Glutathione S-Transferase (GST) is a protein family with multiple physiological functions, primarily located within the cytoplasm. GST is a crucial component of the body's detoxification enzyme system. It mainly catalyzes the covalent binding of the sulfhydryl group of glutathione (GSH) to Glutathione S-Transferase (GST) is a protein family with multiple physiological functions, primarily located within the cytoplasm. GST is a crucial component of the body's detoxification enzyme system. It mainly catalyzes the covalent binding of the sulfhydryl group of glutathione (GSH) to various chemicals and their metabolites, converting electrophilic compounds into hydrophilic compounds that are more easily excreted in bile or urine. This process facilitates the degradation and elimination of potentially toxic substances from the body. Thus, GST plays a vital biological role in protecting cells from damage caused by electrophilic compounds. GST possesses GSH peroxidase activity (also known as non-Se-GSH-Px) and functions in repairing oxidatively damaged macromolecules such as DNA and proteins. The GST-catalyzed reaction consumes GSH but does not increase GSSG levels. Detection Principle: GST catalyzes the conjugation of GSH with CDNB (1-chloro-2,4-dinitrobenzene). The conjugation product has an absorption peak at 340 nm. The GST activity is calculated by measuring the rate of increase in absorbance at 340 nm. Detection Range: 2 - 76 U/L Sensitivity: 2 U/L Applicable Samples: Serum (plasma), animal/plant tissues, cells, bacteriaG1501773Component48T96TStorageG1501773AAssay Buffer60 mL120 mL2-8℃G1501773BChromogen11 mL22 mL2-8℃. Store in the dark.G1501773CSubstrate1EA1EA2-8℃. Store in the dark.Note: Before formal testing, it is recommended to perform a preliminary test with 2-3 samples expected to have significant differences.User-Prepared Instruments and ReagentsMicroplate reader or UV spectrophotometer (capable of measuring absorbance at 340 nm)96-well UV plate or micro quartz cuvettes, adjustable micropipettes and tipsIce maker, refrigerated centrifuge, water bathDeionized waterHomogenizer (for tissue samples)Experimental Procedure1. Reagent PreparationReagent NameReagent PreparationNotesAssay BufferReady-to-use; Equilibrate to room temperature before use.Store at 4℃.ChromogenReady-to-use; Equilibrate to room temperature before use.Store at 4°C protected from light. Skin irritant. Use appropriate protective equipment.Substrate Working ReagentBefore use, dissolve in 2.4 mL of deionized water.Unused reagent can be stored at 4°C protected from light for one month.2. Sample Preparation2.1 Animal/Plant TissuesWeigh 0.1 g of tissue sample. Add 1 mL of pre-cooled Assay Buffer and homogenize quickly on ice. Centrifuge the homogenate at 8,000 g, 4°C for 10 minutes. Collect the supernatant and keep it on ice for assay.2.2 Cells or BacteriaCollect 5×10⁶ cells or bacteria into a centrifuge tube. Wash with cold PBS, centrifuge, and discard the supernatant. Add 1 mL of Assay Buffer. Sonicate in an ice bath for 5 minutes (power 20% or 200W, pulse 3s on, 7s off, repeat 30 times). Centrifuge at 8,000 g, 4°C for 10 minutes. Collect the supernatant and keep it on ice for assay.2.3 Serum (Plasma)Assay directly.Note:Sample processing should be performed on ice. If not used immediately, samples can be stored at -80°C for one month.For GST activity measurement in cells, the cell count should be between 3-5×10⁶. Use Assay Buffer with sonication for cell extraction; do not use cell lysis buffers.If protein concentration measurement is required, Aladdin's BCA Protein Quantification Kit (B665595) or Ready-to-Use BCA Protein Quantification Kit (R1491648) is recommended.3. Assay Steps3.1 Preheat the microplate reader or UV spectrophotometer for at least 30 minutes. Set the wavelength to 340 nm. For UV spectrophotometers, zero the instrument with deionized water.3.2 Incubate the Substrate Working Reagent at 25°C (for general species) or 37°C (for mammals) for 15 minutes.3.3 Add reagents to a 96-well UV plate or micro quartz cuvette as follows:ReagentBlank (µL)Test (µL)Sample020Assay Buffer200Chromogen180180Substrate Working Reagent20203.4 Mix rapidly and immediately measure the change in absorbance at 340 nm. Record the absorbance for the Blank at 10 seconds (A1) and 310 seconds (A2). Record the absorbance for the Test at 10 seconds (A3) and 310 seconds (A4). Calculate ΔA blank = A2 - A1, ΔA test = A4 - A3. Note: Only one Blank is needed. A preliminary test with 2-3 samples showing expected significant differences is recommended. If the sample absorbance is greater than 1, dilute the sample with deionized water and multiply the result by the dilution factor. The reaction temperature significantly affects the results; maintain it at 25°C (general species) or 37°C (mammals). 4. Calculation of Results 4. Calculation of Results Note: We provide both derived and simplified calculation formulas. They are equivalent. The simplified formulas in bold are recommended for final calculation. 4.1 Calculation Formulas for 96-well UV Plate (1) Based on Protein Concentration Unit Definition: One unit of enzyme activity is defined as the amount that catalyzes the conjugation of 1 µmol of CDNB with GSH per minute per milligram of protein at 25°C or 37°C. Calculation Formula: GST Activity (U/mg prot) = (ΔAtest - ΔAblank) ÷ (ε × d) × 10⁶ × Vtotal reaction ÷ (Cpr × Vsample) ÷ T = 0.46 × (ΔAtest - ΔAblank) ÷ Cpr (2) Based on Sample Fresh Weight Unit Definition: One unit of enzyme activity is defined as the amount that catalyzes the conjugation of 1 µmol of CDNB with GSH per minute per gram of sample at 25°C or 37°C. Calculation Formula: GST Activity (U/g fresh weight) = (ΔAtest - ΔAblank) ÷ (ε × d) × 10⁶ × Vtotal reaction ÷ (Vsample ÷ Vtotal sample × W) ÷ T = 0.46 × (ΔAtest - ΔAblank) ÷ W (3) Based on Cell or Bacterial Count Unit Definition: One unit of enzyme activity is defined as the amount that catalyzes the conjugation of 1 µmol of CDNB with GSH per minute per 10⁴ cells or bacteria at 25°C or 37°C. Calculation Formula: GST Activity (U/10⁴) = (ΔAtest - ΔAblank) ÷ (ε × d) × 10⁶ × Vtotal reaction ÷ (500 × Vsample ÷ Vtotal sample) ÷ T = 0.46 × (ΔAtest - ΔAblank) ÷ 500 (4) Based on Liquid Volume Unit Definition: One unit of enzyme activity is defined as the amount that catalyzes the conjugation of 1 µmol of CDNB with GSH per minute per milliliter of liquid at 25°C or 37°C. Calculation Formula: GST Activity (U/mL) = (ΔAtest - ΔAblank) ÷ (ε × d) × 10⁶ × Vtotal reaction ÷ Vsample ÷ T = 0.46 × (ΔAtest - ΔAblank) 4.2 Calculation Formulas for Micro Quartz Cuvette Adjust the pathlength (d) in the formulas above from 0.5 cm to 1 cm for calculations. Parameter Definitions: ε: Molar extinction coefficient of the product, 9.6 × 10³ L/mol/cm d: Light path for 96-well plate (0.5 cm) 10⁶: Conversion factor (1 mol = 1 × 10⁶ µmol) V total reaction : Total reaction volume (220 µL = 2.2 × 10⁻⁴ L) Cpr: Protein concentration of the supernatant (mg/mL) W: Sample weight (g) V sample : Volume of supernatant added to the reaction system (20 µL = 0.02 mL) V total sample : Volume of extraction buffer added (1 mL) T: Reaction time (5 min) 500: Cell or bacterial count factor (5 × 10⁶ total cells / 10⁴ per unit = 500)Precautions1. Biochemical reagents are generally irritating and potentially biologically toxic. For your safety and health, implement appropriate biosafety precautions throughout the experiment, including wearing lab coats, masks, gloves, and head covers. Perform experiments in a fume hood or biosafety cabinet.2. This product is for research use only. Not for use in clinical diagnosis... Read More | The content of this cell is too long for an XLSX file (more than 32767 characters). Please use the CSV format for this export | Products Content:F666101Component500 U5000 UStorageF666101AFastStar Probe Buffer (for bisDNA)2×1.2 mL2×12 mL-20℃. Avoid freeze/thaw cycle. Protect from light.F666101BSuperFastStar DNA Polymerase (5U/µL)100 µL1 mL-20℃. Avoid freeze/thaw cycle. Protect from light.Products Content:F666101Component500 U5000 UStorageF666101AFastStar Probe Buffer (for bisDNA)2×1.2 mL2×12 mL-20℃. Avoid freeze/thaw cycle. Protect from light.F666101BSuperFastStar DNA Polymerase (5U/µL)100 µL1 mL-20℃. Avoid freeze/thaw cycle. Protect from light.Products IntroductionThis product is mainly used for PCR using bisulfite-treated DNA as template, in which SuperFastStar DNA Polymerase is a new high-efficiency hot-start enzyme modified by bis-monoclonal antibody, which is completely blocked at room temperature, thus effectively avoiding non-specific amplification caused by the non-specific binding of the primer to the template or the primer dimerization under the condition of room temperature. The optimized FastStar Probe Buffer (for bisDNA) contains PCR Buffer, dNTPs and Mg2+, etc., which is easy to use as customers only need to add templates, primers and probes.caveat1 Before use, please mix the product gently by turning it up and down after it has been completely melted and centrifuged briefly.2. Avoid repeated freezing and thawing of the product, which may degrade its performance. This product can be stored at -20℃ for a long period of time, protected from light. If frequent use is required within a short period of time, it can be stored at 2-8℃.Usage The following examples are conventional PCR reaction systems and conditions, which should be improved and optimized according to the template, primer structure and target fragment size.1.PCR reaction system Note: 1) Usually, better results can be obtained with a primer concentration of 0.2 µM, and 0.1-1.0 µM can be used as a reference for setting the range.2）The concentration of the probe used is related to the fluorescence quantitative PCR instrument used, the type of probe, and the type of fluorescent labeling substance, so please refer to the instrument manual or the specific requirements for the use of each fluorescent probe to adjust the concentration.3）Usually the amount of DNA template is 10-100 ng of genomic DNA or 1-10 ng of cDNA as a reference. Since the templates of different species contain different copy numbers of the target gene, the templates can be diluted in gradients to determine the optimal amount of template to use.2.PCR reaction conditionsNote: 1) The initial denaturation of this product at 95°C for 30s is sufficient for enzyme activation; complex templates can be extended to 3min denaturation.(2) It is recommended to use two-step PCR reaction program, if you can't get good experimental results due to the use of primers with lower Tm value, etc., you can try three-step PCR amplification, and the annealing temperature should be set in the range of 56℃-64℃ as a reference... Read More | RAFT Agent Kit for controlling polymerizations at the molecular level detailed list of products: Catalog Number Product Name Component Catalog Number Component Name Component CAS Specification&Purity R396714 RAFT Agent Kit for controlling polymerizations at the molecular level C139356-500mg 4-RAFT Agent Kit for controlling polymerizations at the molecular level detailed list of products: Catalog Number Product Name Component Catalog Number Component Name Component CAS Specification&Purity R396714 RAFT Agent Kit for controlling polymerizations at the molecular level C139356-500mg 4-Cyano-4-(dodecylsulfanylthiocarbonyl)sulfanylpentanoic acid 870196-80-8 See Component Catalog Number R396714 RAFT Agent Kit for controlling polymerizations at the molecular level C396701-500mg Cyanomethyl dodecyl trithiocarbonate 796045-97-1 See Component Catalog Number R396714 RAFT Agent Kit for controlling polymerizations at the molecular level C396703-500mg Cyanomethyl methyl(phenyl)carbamodithioate 76926-16-4 See Component Catalog Number R396714 RAFT Agent Kit for controlling polymerizations at the molecular level C168358-500mg 2-Cyano-2-propyl benzodithioate 201611-85-0 See Component Catalog Number R396714 RAFT Agent Kit for controlling polymerizations at the molecular level C396706-500mg 2-(2-Cyanoprop-2-yl)-S-dodecyltrithiocarbonate 870196-83-1 See Component Catalog Number R396714 RAFT Agent Kit for controlling polymerizations at the molecular level C132316-500mg 4-Cyano-4-(phenylcarbonothioylthio)pentanoic Acid 201611-92-9 See Component Catalog Number R396714 RAFT Agent Kit for controlling polymerizations at the molecular level S396708-500mg S,S-Dibenzyl trithiocarbonate 26504-29-0 See Component Catalog Number R396714 RAFT Agent Kit for controlling polymerizations at the molecular level D396711-500mg 2-(Dodecylthiocarbonothioylthio)-2-methylpropionic acid 461642-78-4 See Component Catalog Number... Read More | This reagent kit is based on TRIzon's improved columnar total RNA extraction kit. This product can be extracted from animal groupsExtract total RNA from samples such as textiles, plant materials, various microorganisms, and cultured cells. Firstly, the cracking solution is fully cracked This reagent kit is based on TRIzon's improved columnar total RNA extraction kit. This product can be extracted from animal groupsExtract total RNA from samples such as textiles, plant materials, various microorganisms, and cultured cells. Firstly, the cracking solution is fully cracked andHomogenized samples, in their unique high salt state, RNA specifically binds to silicon matrix membranes, greatly reducingEffectively removing organic solvent contamination while removing protein contamination, resulting in higher purity and quality of RNA. bookThe product can quickly extract total RNA from various cells or tissues, and can process 30-50 mg of tissue or 5 × 10 ⁶ cells each time,Can handle multiple different samples simultaneously. If it is an RNA experiment that is very sensitive to trace amounts of DNA, the residual DNA can be utilizedUsing DNase without RNase for digestion and removal on the column, the extracted RNA can be directly applied to RT-PCR Experiments such as Northern Blot, Dot Blot, and in vitro translation. U665516 Component 50 T Storage U665516A DNase I 1000 U -20℃. Avoid freeze/thaw cycle. U665516B 10×Reaction Buffer 1000 µL -20℃. Avoid freeze/thaw cycle. U665516C TRIzon Reagent 60 mL 2-8℃. Protect from light. U665516D TRIzon PaI™ 10 mL 2-8℃. Protect from light. U665516E Buffer RW1 40 mL RT U665516F Buffer RW2 (concentrate) 11 mL RT U665516G RNase-Free Water 10 mL RT U665516H Spin Columns RM with Collection Tubes 50 sets RT U665516I RNase-Free Centrifuge Tubes (1.5 mL) 50 EA RTPreparation and important precautions before the experiment:1.To prevent RNase pollution, attention should be paid to the following aspects:1) RNase's plastic products and gun heads to avoid cross contamination.2) Prepare the solution using water without RNase.3) Operators should wear disposable masks and gloves, and change gloves frequently during the experiment.2. The sample should avoid repeated freezing and thawing, otherwise it will affect the yield and quality of RNA extraction.3. If TRIzon Reagent is found to have precipitates before use, it can be dissolved in a water bath at 56 ℃ for a few minutes.Before the first use, anhydrous ethanol should be added to Buffer RW2 according to the instructions on the reagent bottle label.5. All centrifugation steps should be carried out at room temperature unless otherwise specified, and all operation steps should be carried out quickly.Usage：1. Sample processing1a. Organization: 30-50 mg of tissue is thoroughly ground in liquid nitrogen and 1 mL of TRIzon Reagent is added, or 1 mL of TRIzon Reagent is added to the tissue sample and homogenized. Attention: The sample volume should not exceed 10% of the volume of TRIzon Reagent.2a. Single layer cell culture: Remove the culture medium and add an appropriate amount every 10 cm ² Add 1 mL of TRIzon Reagent.3a. Cell suspension: Collect cells by centrifugation. Add 1 mL of TRIzon Reagent to every 5 × 10 µ m cell.2. After adding TRIzon Reagent, repeatedly blow a few times to fully crack the sample. Leave at room temperature for 5 minutes to completely separate the protein nucleic acid complex.3. Add 200 to every 1 mL of TRIzon Reagent µ LTRIzon PaI ™， Cover the tube tightly, vigorously shake for 15 seconds, and let it sit at room temperature for 2 minutes.4. Centrifuge at 4 ℃ 12000 rpm (~13400 × g) for 10 minutes. At this time, the sample is divided into three layers: the red organic phase, the middle layer, and the upper colorless aqueous phase. RNA is mainly in the upper aqueous phase. Move the upper aqueous phase to a new RNase Free centrifuge tube (provided).5. Add an equal volume of 70% ethanol (prepared without RNase water) to the obtained aqueous solution, invert and mix well.6. Add all the solutions obtained in the previous step to the spin columns RM that have been loaded into the collection tube. If the solution cannot be added at once, it can be transferred in multiple batches. Centrifuge at 12000 rpm for 20 seconds, discard the waste liquid in the collection tube, and place the adsorption column back into the collection tube.7. Add 350 to the adsorption column µ L Buffer RW1, centrifuge at 12000 rpm for 20 seconds, discard the waste liquid in the collection tube, and place the adsorption column back into the collection tube.8. Preparation of DNase I mixture: Take 52 µ LRNase Free Water, add 8 to it µ L 10 x Reaction Buffer and 20 µ L DNase I (1 U/ µ L) Mix well and prepare to a final volume of 80 µ The reaction solution of L.9. Directly add 80 µ L DNase I mixture to the adsorption column and incubate at 20-30 ℃ for 15 minutes.10. Add 350 to the adsorption column µ L Buffer RW1, centrifuge at 12000 rpm for 1 minute, discard the waste liquid, and place the adsorption column back into the recovery manifold.11. Add 500 to the adsorption column µ L Buffer RW2 (check if anhydrous ethanol has been added before use), centrifuge at 12000 rpm for 20 seconds, discard the waste liquid in the collection tube, and place the adsorption column back into the collection tube.12. Repeat step 11.Centrifuge at 12000 rpm for 2 minutes and discard the waste liquid from the collection tube. Place the adsorption column at room temperature for a few minutes and thoroughly air dry. Attention: The purpose of this step is to remove residual ethanol from the adsorption column, which will affect subsequent enzymatic reactions (enzyme digestion,. )PCR, etc.14. Place the adsorption column in a new RNase free centrifuge tube and add 30-50 to the middle of the adsorption column µ Place RNase Free Water at room temperature for 1 minute, centrifuge at 12000 rpm for 1 minute, collect RNA solution, and store RNA at -70 ℃ to prevent degradation.Attention:1) The volume of RNase Free Water should not be less than 30 µ L. Small volume affects the recovery rate.2) If you want to increase RNA production, you can use 30-50 µ Repeat step 14 for the new RNase Free Water.3) If you want to increase the RNA concentration, you can add the obtained solution back to the adsorption column and repeat step 14... Read More |