| Description | Reducing sugars (RS) are widely present in animals, plants, microorganisms, and cultured cells. Reducing sugars in plants primarily include glucose, fructose, and maltose. Among these, glucose and fructose are not only the main substrates for respiration but also serve as substrates for the further Reducing sugars (RS) are widely present in animals, plants, microorganisms, and cultured cells. Reducing sugars in plants primarily include glucose, fructose, and maltose. Among these, glucose and fructose are not only the main substrates for respiration but also serve as substrates for the further synthesis of sucrose, starch, and cellulose.Detection Principle: In an alkaline solution, 3,5-dinitrosalicylic acid (DNS) can be reduced by reducing sugars to produce a brown-red-colored amino compound, which has a characteristic absorption peak at 540 nm. Within a certain concentration range, the RS content is linearly correlated with the absorbance at 540 nm. The RS content in the sample can be calculated based on a standard curve.Detection Range: 0.05 - 0.6 mg/mLSensitivity: 0.025 mg/mLApplicable Samples: Plant tissues, animal tissues, cells, bacteria, serum (plasma)R1501790Component48T96TStorageR1501790AExtraction Buffer60 mL120 mL2-8℃R1501790BDNS Reagent10 mL20 mL2-8℃. Store in the dark.R1501790CStandard1EA1EA2-8℃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 visible spectrophotometer (capable of measuring absorbance at 540 nm)96-well plate or micro glass cuvettes, adjustable micropipettes and tipsCentrifuge, water bathDeionized waterHomogenizer (for tissue samples)Experimental Procedure1. Reagent PreparationReagent NameReagent PreparationNotesExtraction BufferReady-to-use; Equilibrate to room temperature before use.Store at 4°C. Slightly irritating. Use appropriate personal protective equipment.DNS ReagentReady-to-use; Equilibrate to room temperature before use.Store at 4°C protected from light. Slightly irritating. Use appropriate personal protective equipment.StandardBefore use, add 1 mL of deionized water to dissolve, preparing a 10 mg/mL stock standard solution.Can be stored at 4°C for 2 weeks.2. Standard Curve SetupDilute the 10 mg/mL standard stock solution with deionized water to concentrations of 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, and 0.05 mg/mL.TubeVolume of 10 mg/mL Standard (µL)Volume of Deionized Water (µL)Concentration (mg/mL)Std.1609400.6Std.2509500.5Std.3409600.4Std.4309700.3Std.5209800.2Std.6109900.1Std.759950.05Note: The standard curve must be generated with each experiment. Diluted standard solutions are unstable and must be used within 4 hours.3. Sample Preparation3.1 Plant or Animal Tissue SamplesWeigh approximately 0.1 g of tissue. Add 1 mL of Extraction Buffer and homogenize in an ice bath. Transfer the homogenate to a capped centrifuge tube (to prevent evaporation during heating). Incubate in an 80°C water bath for 40 minutes, vortexing every 5 minutes. Centrifuge at 8,000 g, 25°C for 10 minutes. Collect the supernatant for assay.3.2 Bacteria or CellsCollect bacteria or cells into a centrifuge tube; discard the supernatant. Add 1 mL of Extraction Buffer per 5 million bacteria/cells. Sonicate in an ice bath for 5 minutes (power 20%, pulse 3s on, 10s off, repeat 30 times). Transfer to a capped centrifuge tube (to prevent evaporation during heating). Incubate in an 80°C water bath for 40 minutes, vortexing every 5 minutes. Centrifuge at 8,000 g, 25°C for 10 minutes. Collect the supernatant for assay.3.3 Serum (Plasma) SamplesTake 0.1 mL of serum (plasma) and add 0.9 mL of Extraction Buffer; mix thoroughly. Transfer to a capped centrifuge tube (to prevent evaporation during heating). Incubate in an 80°C water bath for 40 minutes, vortexing every 5 minutes. Centrifuge at 8,000 g, 25°C for 10 minutes. Collect the supernatant for assay.Note:If protein concentration measurement is required, Aladdin's BCA Protein Quantification Kit (B665595) or Ready-to-Use BCA Protein Quantification Kit (R1491648) is recommended. The Extraction Buffer contains components that denature proteins. If calculating based on protein concentration, protein needs to be re-extracted separately for measurement.4. Assay Steps4.1 Preheat the microplate reader or visible spectrophotometer for at least 30 minutes. Set the wavelength to 540 nm. For spectrophotometers, zero the instrument with deionized water.4.2 Assay Procedure:ReagentBlank Tube (µL)Standard Tube (µL)Test Tube (µL)Control Tube (µL)Sample00175175Standard (various conc.)017500Deionized Water17500125DNS Reagent1251251250Mix well. Heat in a boiling water bath for 5 minutes (cap tightly to prevent evaporation). Remove and immediately cool to room temperature. Transfer 200 µL to a 96-well plate or micro glass cuvette. Measure the absorbance at 540 nm. Calculate ΔA test = A test - A control, ΔA standard = A standard - A blank. Note:The Blank and Standard tubes only need to be set up 1-2 times.It is recommended to perform a preliminary test with 2-3 samples expected to have significant differences before the formal experiment. If ΔA <sub> test </sub> is less than 0.04, consider increasing the sample volume appropriately. If ΔA <sub> test </sub> is greater than the ΔA <sub> standard </sub> of the 0.6 mg/mL standard, further dilute the sample with Extraction Buffer (multiply the result by the dilution factor) or reduce the amount of sample used for extraction.5. Calculation of ResultsNote: We provide both the derived formula and a simplified formula. They are equivalent. It is recommended to use the simplified formula in bold for final calculation.5.1 Standard Curve PlottingPlot the standard concentration (y-axis) against ΔA standard (x-axis) to generate the standard curve. Substitute ΔA test into the standard curve equation to calculate y (mg/mL).5.2 Sample Reducing Sugar Content Calculation(1) Based on Sample WeightReducing Sugar (µg/g) = 1000 × y × V<sub>extraction</sub> ÷ W × n = 1000 × y / W × n(2) Based on Sample Protein ConcentrationReducing Sugar (µg/mg prot) =1000 × y × Vextraction ÷ (Vextraction × Cpr) × n=1000 × y / Cpr × n(3) Based on Bacterial or Cell CountReducing Sugar (µg/10⁴) =1000 × y × V<sub>extraction</sub> ÷ 500 × n = 2 × y × n(4) Based on Serum (Plasma) VolumeReducing Sugar (µg/mL) = 1000 × y × Vextraction ÷ Vliquid × n = 10000 × y × nParameter Definitions:1000: Unit conversion factor (1 mg/mL = 1000 µg/mL)V extraction : Volume of Extraction Buffer added (1 mL)V liquid : Volume of serum (plasma) added (0.1 mL)Cpr: Sample protein concentration (mg/mL)W: Sample weight (g)500: Total number of bacteria or cells (5 million)n: Dilution factor6. Representative ResultsTypical Standard Curve: y = 0.2243x + 0.0545, R² = 0.9957 PrecautionsThis product is for research use only. Not for use in clinical diagnosis. For your safety and health, please wear lab coats and disposable gloves during operation... Read More | CFDASE cell proliferation and tracking detection kit is a kit for cell proliferation and tracking detection based on CFDA se. This kit is composed of CFDASE powder, solvent and staining buffer. CFDASE is a derivative of fluorescein diacetate (FDA), which has cell membrane permeability and CFDASE cell proliferation and tracking detection kit is a kit for cell proliferation and tracking detection based on CFDA se. This kit is composed of CFDASE powder, solvent and staining buffer. CFDASE is a derivative of fluorescein diacetate (FDA), which has cell membrane permeability and does not have fluorescence luminescence. When CFDASE penetrates the cell membrane into living cells, it can be catalysed by esterases in the cytosol to produce carboxyfluorescein succinimidyl ester (CFSE), which can emit strong green fluorescence, cannot penetrate the cell membrane, and can remain intact in the cell. CFSE can also spontaneously and irreversibly covalently bind to intracellular amino groups to couple to cellular proteins. Meanwhile, the excess and uncoupled CFDASE returned to the extracellular medium by passive diffusion and was cleared by subsequent washing steps. The fluorescence of non dividing cells labeled by CFDASE is very stable, and the stable labeling time can reach several months, so it is very suitable for cell community analysis. The fluorescence of CFDASE labeled cells is very homogeneous, which is superior to other cell tracking fluorescent probes used previously, such as PKH26, and the fluorescence distribution of the divided progeny cells is also very uniform. In the process of cell division and proliferation, CFSE labeled fluorescence can be evenly distributed to the two progeny cells, and the fluorescence intensity becomes half of the parental cells. According to the fluorescence intensity, flow cytometer (FL1 channel) can detect undivided cells, cells that divide once (1 / 2 of the fluorescence intensity), twice (1 / 4 of the fluorescence intensity), three times (1 / 8 of the fluorescence intensity), and cells that divide more times. CFDASE can detect up to eight or more cleavages. CFDASE labeled cells can be used for proliferation studies in vitro and in vivo, and have the function of not staining adjacent cells. CFDASE is most commonly used to detect the proliferation of lymphocytes, and can also be used to detect the proliferation of fibroblasts, NK cells and other cells. CFDASE labeled cells showed green fluorescence. In addition to flow cytometry to detect cell proliferation, fluorescence microscopy can also be used for homogeneous staining of cell tracking observation.Components:ComponentsC598182-20TC598182-500TA. CFDA SE1 tube1 tubex5B.CFDA SE solvent20 µL500 µLC.10x CFDA SE Buffer1 mL x250 mLMatters needing attention：1. please centrifuge the product to the bottom of the tube immediately before use, and then conduct subsequent experiments. 2. CFDA and Se are easily hydrolyzed and will deteriorate quickly in aqueous solution. Please avoid contact with water during use. Contact with water during the process of labeling cells is within the permitted range. 3. CFDA se solvent will solidify at lower temperatures such as 4 º C and ice bath and stick to the bottom, wall or cover of the centrifugal tube. It can be used after incubating in a 20-25 º C water bath for a while until it is completely dissolved. 4. this kit optimizes the CFDA se staining system, but users are advised to explore the optimal working concentration and staining time according to their own cell type, culture conditions and application direction. Different cells have different lactonase activities, so the staining effect is different. 5. fluorescent dyes have quenching problems. Please avoid light during operation to slow down fluorescence quenching. 6. for your safety and health, please wear experimental clothes and disposable gloves.Usage method:1. Preparation of reagents(1) Preparation of CFDA SE storage solution: Take one tube of CFDA SE provided in the reagent kit and restore it to room temperature. Instantly centrifuge to allow the powder to fully settle to the bottom of the tube. Add 100 µ L CFDA SE solvent (add 20 µ L CFDA SE solvent) to it and dissolve it thoroughly to prepare CFDA SE storage solution (1000 ×). Prepared CFDA SE storage solution, stored at -20 ℃ in the dark, with a shelf life of two months- Storing at 70 ℃ in the dark can extend the usage time appropriately.(2) Preparation of CFDA SE Buffer: Dilute 10 x CFDA SE Buffer to 1 x with sterile cell culture grade water as needed. The prepared 1 × CFDA SE Buffer can be stored at 4 ℃ and can be stored at -20 ℃ if not in use for a long time.2. Marking and detection(1) Centrifuge the collected cells, use 1 mL 1 × CFDA SE Buffer to re suspend the cells in a 15 mL centrifuge tube, and adjust the cell concentration to 1-5 × 106 cells/mL.(2) Preparation of CFDA SE working solution: Dilute the CFDA SE storage solution (1000 ×) with 1 × CFDA SE Buffer to 2 ×.(3) Staining: Add 1 mL of CFDA SE working solution (2 x) to 1 mL of cell suspension to be labeled, invert and mix well, and incubate at 37 ℃ for 10 minutes.(4) Immediately add 5 times the volume of preheated complete culture medium (including serum) to the centrifuge tube, invert and mix well to terminate the labeling reaction.(5) Centrifuge at 1000 rpm for 5 minutes at room temperature to remove the supernatant, then wash once with 5-10 mL of complete culture medium.(6) Add 5-10 mL of complete culture medium and incubate at 37 ℃ for 5 minutes to promote the residence of CFDA SE in the cells and the entry of unreacted CFDA SE into the complete cell culture medium. Centrifuge at 1000 rpm for 5 minutes at room temperature to remove the supernatant and complete the final wash.(7) Subsequently, the cells can be cultured using the normal cultivation method. The labeling effect can be directly observed under a fluorescence microscope, or cell proliferation can be detected by flow cytometry after appropriate cultivation time, showing green fluorescence. The labeled cells can also be used for transplantation in live animals and for fluorescence tracing.Note: a If cell fixation is required, use aldehyde fixatives such as 4% paraformaldehyde to fix at room temperature for 15 minutes; If additional labeling such as antibody labeling is required afterwards, please permeabilize the cells with ice acetone for 10 minutes. b. The optimal labeling concentration and incubation time for CFDA SE vary for different cells. The initial experiment can be conducted according to the experimental steps. If the effect is not satisfactory, it is recommended to adjust the staining concentration and incubation time to achieve the best labeling effect.Scope of application：Cell proliferation assay... Read More | Inquire | Products contentN665989Component240 TStorageN665989AIndex N501 Primers for Illumina240 µL-20℃. Avoid freeze/ Thaw cycle.N665989BlIndex N901-N924 Primers for Illumina24×10 µL-20℃. Avoid freeze/ Thaw cycle.Note: The amount of individual primers used is 1 µl, each N7-endProducts contentN665989Component240 TStorageN665989AIndex N501 Primers for Illumina240 µL-20℃. Avoid freeze/ Thaw cycle.N665989BlIndex N901-N924 Primers for Illumina24×10 µL-20℃. Avoid freeze/ Thaw cycle.Note: The amount of individual primers used is 1 µl, each N7-end primer can perform 10 DNA library constructs, and each kit can perform 240 DNA library constructs. Products IntroductionThis kit is a companion kit to the transposase-based Rapid DNA Library Construction Kit for Illumina platform library construction. Each kit contains one N5 primer and 24 N7 primers, which can be used to prepare 24 different single-ended Index libraries. All reagents provided in the kits have been subjected to stringent quality control and functional validation to maximize the stability and reproducibility of library construction. The libraries can be used for sequencing on Illumina platforms such as HiSeq X-10/4000/2500/2000 and MiSeq. Provide your own instruments, reagents and consumables1. Magnetic frame: DynaMagTM-2 is recommended.2. DNA purification and recovery kit: It is recommended to use Kangwei DNA purification and recovery kit by magnetic bead method.3. DNA building kit: It is recommended to use the Kangwei Century transposase method second-generation sequencing rapid DNA building kit.4. Anhydrous ethanol.5. Reaction tubes: It is recommended to use low adsorption PCR tubes with 1.5 ml centrifuge tubes;Tip: It is recommended to use a high quality filter tip to prevent contamination of kits and library samples. Pre-experiment Preparation and Important NotesPlease centrifuge briefly before opening the cap so that the liquid collects at the bottom of the tube to avoid cross-contamination between different primers. ProcedureFor the use of the CombiVision Second Generation Sequencing Multisample Primer Kit, please follow the CombiVision Second Generation Sequencing Rapid DNA Library Kit protocol.Index N501 Primer for IlluminaIndex N901-N996 Primer for Illumina... Read More | Products contentN665993Component240 TStorageN665993AIndex N501 Primers for Illumina240 µL-20℃. Avoid freeze/ Thaw cycle.N665993BIndex N925-N948 Primers for Illumina24×10 µL-20℃. Avoid freeze/ Thaw cycle. Products Introduction This kit is a companion kit to the transposase-Products contentN665993Component240 TStorageN665993AIndex N501 Primers for Illumina240 µL-20℃. Avoid freeze/ Thaw cycle.N665993BIndex N925-N948 Primers for Illumina24×10 µL-20℃. Avoid freeze/ Thaw cycle. Products Introduction This kit is a companion kit to the transposase-based Rapid DNA Library Construction Kit for Illumina platform library construction. Each kit contains one N5 primer and 24 N7 primers, which can be used to prepare 24 different single-ended Index libraries. All reagents provided in the kits have been subjected to stringent quality control and functional validation to maximize the stability and reproducibility of library construction. The libraries can be used for sequencing on Illumina platforms such as HiSeq X-10/4000/2500/2000 and MiSeq. Provide your own instruments, reagents and consumables1. Magnetic frame: DynaMagTM-2 is recommended.2. DNA purification and recovery kit: It is recommended to use Kangwei DNA purification and recovery kit by magnetic bead method.3. DNA building kit: It is recommended to use the Kangwei Century transposase method second-generation sequencing rapid DNA building kit.4. Anhydrous ethanol.5. Reaction tubes: It is recommended to use low adsorption PCR tubes with 1.5 ml centrifuge tubes;Tip: It is recommended to use a high quality filter tip to prevent contamination of kits and library samples. Pre-experiment Preparation and Important NotesPlease centrifuge briefly before opening the cap so that the liquid collects at the bottom of the tube to avoid cross-contamination between different primers. ProcedureFor the use of the CombiVision Second Generation Sequencing Multisample Primer Kit, please follow the CombiVision Second Generation Sequencing Rapid DNA Library Kit protocol.Index N501 Primer for IlluminaIndex N901-N996 Primer for Illumina... Read More |