Lumefantrine D18_1185240-53-2_DataSheet_MedChemExpress

Endotoxin Removal Solution Catalog Number E4274Product DescriptionEndotoxins are lipopolysaccharides (LPS), a major component of the Gram-negative bacterial cell wall, and are commonly found as contaminants in plasmid DNA preparations from E. coli. Endotoxins are large, negatively charged molecules that co-purify with DNA on ion exchange and size exclusion columns and in CsCl banding. Endotoxins are extremely potent stimulators of the mammalian immune system and are toxic to primary cells and to animals. The endotoxin toxicity is an obstacle to in vitro and in vivo transfection experiments.Non-ionic detergents, traditionally used for separation of integral membrane proteins,1 can be utilized for removal of endotoxins from DNA solutions by phase separation.2The solubility behavior of a detergent in a dilute, aqueous solution at physiological salt and pH conditions is strongly dependent upon the temperature of the solution. At low temperatures, the detergent forms a clear, micellar solution, but above the cloud point temperature, the micelles form larger, turbid aggregates and ultimately fuse to form a separate phase. The lower phase is detergent-enriched and the detergent-depleted upper phase contains detergent at a concentration slightly above the critical micellar concentration (CMC). Amphiphilic and hydrophobic molecules associated with the micelles of the detergent will aggregate within the detergent-enriched phase, while the soluble, hydrophilic molecules will remain in the detergent-depleted upper phase.Extraction of endotoxin contaminated DNA solutions with the appropriate non-ionic detergent will separate the hydrophilic DNA from the amphiphilic endotoxin. The amphiphilic endotoxin will associate with the lower phase, while the DNA will remain in the upper, detergent-depleted phase.2Reagents and equipment required, but not provided • Water, Molecular Biology Reagent, Catalog Number W4502• E-TOXATE® Water, Catalog Number 2107, or Tris-EDTA (TE) buffer 100×, Catalog NumberT9285• DNA solution (0.5 ml), ~ 1 mg/ml in E-TOXATE®Water or TE buffer• 3 M sodium acetate solution, pH 7.5.• 2-Propanol, Catalog Number I9516, or Ethanol, 190 proof, Catalog Number E7148; 200 proof, CatalogNumber E7023• 70% Ethanol• E-TOXATE®reagents Kits, Catalog Numbers 210A1, 210B1 or 210C1• Ice bucket• Heat block or incubator at 37 °C• Microcentrifuge at room temperature• 1.5 or 2 ml sterile microcentrifuge tubes• Endotoxin-free pipet tips (40-200 µl, 200-1000 µl) Precautions and DisclaimerThis product is for R&D use only, not for drug, household, or other uses. Please consult the Material Safety Data Sheet for information regarding hazards and safe handling practices.StorageStore at room temperature.Note: Removal of endotoxins from DNA preparations can be performed either during the final stage of DNApreparation, or during an earlier stage.Procedures for Endotoxin RemovalDuring the final stage of DNA preparationNote: The procedure described below was performed on plasmid DNA produced in E. coli DH5α cells.• Losses of up to 50% of the DNA are expected. • Use of a DNA concentration above therecommended 1 mg/ml reduces the efficiency ofthe procedure.1. Pipette 500 µl of the DNA solution into a sterilemicrocentrifuge tube.2. Add 50 µl of the 3 M sodium acetate solution to theDNA sample.3. Incubate on ice for 5 minutes.4. Add 100 µl of cold Endotoxin Removal Solution.5. Mix thoroughly and incubate on ice for 10 minutes.The solution should be light blue and clear.6. Incubate the tube at 37 °C for 20 to 30 minutes oruntil the phases separate.7. Spin for 5 minutes at 3000 x g in themicrocentrifuge. The upper phase is colorless and clear, while the lower phase is blue.8. Carefully transfer the upper phase containing theDNA to a clean microcentrifuge tube.9. Repeat steps 4 through 8 twice.10. Add 0.6× volume of 2-propanol. Mix by inversion atroom temperature and centrifuge at 15,000 x g for30 minutes at 4 °C. Alternatively, add2.5× volumes of ethanol. Incubate overnight at –20°C or 20 minutes at –70 °C and centrifuge at15,000 x g for 30 minutes at 4 °C.11. Carefully remove the supernatant12. Wash the DNA pellet twice with cold 70% ethanol.Remove the supernatant.13. Air-dry the pellet.14. Suspend the DNA in 100 µl of endotoxin free wateror TE buffer.15. Determine DNA concentration and endotoxin levelsusing endotoxin assay reagents and compare tothe starting material. During an earlier stage of DNA preparationThis procedure is based on the alkaline lysis of E. coli DH5α cells.3 The endotoxins are removed immediately after alkaline cell lysis, neutralization, and a clarification step. The resulting high salt solution is suitable for the endotoxin removal step. It is performed under “endotoxin free” conditions. The plasticware used is either sterile and disposable, or NaOH-treated. The buffers are prepared with endotoxin free water.1. Add the Endotoxin Removal Solution (0.2× volume)to the cold, crude DNA solution.2. Incubate on ice and mix occasionally by inversionto obtain a homogenous, clear blue solution3. Incubate at 37 °C for 20 to 30 minutes until thephase separation is obvious.4. Spin for 5 minutes at low speed (3000 x g) at roomtemperature.5. Transfer the upper aqueous phase to an endotoxinfree container.6. Proceed with the DNA purification by any method.Use endotoxin-free buffers and containers. References1. Bordier, C., J. Biol. Chem., 256, 1604-1607, (1981).2. Cotten, M. et al., Gene Therapy, 1, 239-246,(1994).3. Sambrook et al., Molecular Cloning, a LaboratoryManual, 2nd Ed. p. 1.38RK,PHC 09/05-1Sigma brand products are sold through Sigma-Aldrich, Inc.Sigma-Aldrich, Inc. warrants that its products conform to the information contained in this and other Sigma-Aldrich publications. Purchaser must determine the suitability of the product(s) for their particular use. Additional terms and conditions may apply. Please see reverse side ofthe invoice or packing slip.。

HANKS' BALANCED SALTS [HBSS]Without calcium chloride, magnesium sulfate and sodium bicarbonate Product Number H2387Product DescriptionAlthough there have been many modifications to the original formulas in efforts to produce fully defined media, salt solutions still play an important role in tissue culture. A salt solution's basic function, to maintain the pH and osmotic balance in the medium and to provide the cells with water and essentialinorganic ions, is as valuable today as when it was first developed a century ago.Componentsg/L Potassium Chloride0.4Potassium Phosphate Monobasic 0.06(anhydrous)Sodium Chloride8.0Sodium Phosphate Dibasic(anhydrous)0.04788D -Glucose1.0Phenol Red•Na0.011Precautions and Disclaimer REAGENTFor R&D use only. Not for drug, household or other uses.Preparation InstructionsPowdered salts are hygroscopic and should beprotected from moisture. The entire contents of each package should be used immediately after opening. Preparing a concentrated salt solution is notrecommended as precipitates may form. Supplements can be added prior to filtration or introduced aseptically to sterile salt solution.1.Measure out 90% of final required volume ofwater. Water temperature should be 15-20 ˚C.2.While gently stirring the water, add the powderedmedium. Stir until dissolved. Do NOT heat. 3.Rinse original package with a small amount ofwater to remove all traces of powder. Add to solution in step 2.4.To the solution in step 3, add 0.35 g sodiumbicarbonate or 4.7 ml of sodium bicarbonatesolution [7.5%w/v] for each liter of final volume of medium being prepared. Stir until dissolved.5.While stirring, adjust the pH of the medium to 0.1-0.3 pH units below the desired pH since it may rise during filtration. The use of 1N HCl or 1N NaOH is recommended.6.Add additional water to bring the solution to finalvolume.7.Sterilize immediately by filtration using amembrane with a porosity of 0.22 microns.8.Aseptically dispense medium into sterile container. Storage and StabilityStore the dry powdered salts at 2-8 °C under dry conditions and liquid medium at 2-8 °C in the dark. Deterioration of the powdered medium may be recognized by any or all of the following: [1] color change, [2] granulation/clumping, [3] insolubility.Deterioration of the liquid medium may be recognized by any or all of the following: [1] pH change, [2] precipitate or particulates, [3] cloudy appearance [4] color change. The nature of supplements added may affect storage conditions and shelf life of the medium. Product label bears expiration date.ProcedureMaterials Required but Not Provided:Water for tissue culture [W3500]Sodium Bicarbonate [S5761] orSodium Bicarbonate Solution, 7.5% [S8761]1N Hydrochloric Acid [H9892]1N Sodium Hydroxide [S2770]Medium additives as requiredReference1.Hanks, J. (1976) Hanks' Balanced SaltSolution and pH Control. Tissue Culture Association Manual. 3, 3.Revised: March 2007Sigma-Aldrich, I nc. warrants that its products conform to the information contained in this and other Sigma-Aldrich publications. Purchaser must determine the suitability of the product(s) for their particular use. Additional terms and conditions may apply. Please see reverse side of the invoice or packing slip.。

AccuMelt™ HRM SuperMixCat No. 95103-250Size: 250 x 20-µL reactions (2 x 1.25 mL)Store at -25ºC to - 15°C 95103-0121250 x 20-µL reactions (10 x 1.25 mL)protected from light DescriptionAccuMelt HRM SuperMix is a 2X concentrated, ready-to-use reaction cocktail for detection of genetic variations using high resolution melting (HRM) analysis. It includes all required components except for primers and DNA template. HRM is a closed tube, rapid and cost effective procedure for characterization of sequence differences immediately following PCR amplification. It is based on the melting (dissociation) behavior of a PCR product as it transitions from double-stranded to single-stranded DNA in the presence of a fluorescent dsDNA-binding dye. The melting properties of a given PCR product are dependent upon the base composition, length, and strand base-pairing. HRM analysis tools exploit differences in melt curve shapes and DNA melting temperature (Tm) to discriminate sequence differences between samples.AccuMelt HRM SuperMix contains the green-fluorescent dye SYTO® 9 in a stabilized master mix that eliminates the need for time consuming optimization of critical PCR components. This dye provides strong fluorescent signal upon binding to dsDNA at saturating concentrations without inhibiting PCR. The unique chemical composition of this SuperMix further enhances and maximizes the impact of sequence variations on melt curve behavior. This facilitates discrimination of all sequence variations including difficult to resolve base-neutral transversions such as A>T class 4 SNPs. The dNTP mix in AccuMelt HRM SuperMix includes an optimized blend of dTTP and dUTP. This feature supports the optional use of uracil-DNA glycosylase (UNG) to prevent amplification of carry-over contamination, while providing high product yield and reliable PCR performance.Highly specific amplification with high product yield from complex genomic DNA template is critical for successful HRM studies. A key component of this SuperMix is AccuStart™ Taq DNA polymerase, which contains monoclonal antibodies that bind to the polymerase and keep it inactive prior to the initial PCR denaturation step. Upon heat activation at 95ºC, the antibodies denature irreversibly, releasing fully active, unmodified Taq DNA polymerase. This enables specific and efficient primer extension with the convenience of room temperature reaction assembly. AccuMelt HRM SuperMix can be used with all currently available HRM analysis systems. For HRM applications and more detailed product information, please visit our web site at .ComponentsAccuMelt HRM SuperMix (2X): 2X reaction buffer containing optimized concentrations of MgCl2, dNTPs(including dUTP), AccuStart Taq DNA Polymerase, SYTO 9 green-fluorescentdye, and stabilizers.[free Mg ++] = 0.8 mM at 1X final concentrationStorage and StabilityStore components in a constant temperature freezer at -25°C to -15°C protected from light upon receipt.For lot specific expiry date, refer to package label, Certificate of Analysis or Product Specification Form.Guidelines for PCR amplification and HRM analysis:The design of highly specific primers is the single most important parameter for successful PCR amplification and HRM analysis. The use of computer aided primer design programs is encouraged in order to minimize the potential for internal secondary structure and complementation at 3’-ends within each primer and the primer pair. Primer T m should be between 56 to 63ºC and the T m difference between the forward and reverse primers should be less than 2ºC.Amplicon size should be less than 250 bp. Smaller amplicons (60 to 100 bp) generally facilitate HRM discrimination of homozygote samples. We recommend designing and evaluating multiple primer set designs for any given application.Optimal primer concentration may vary between 100 and 500 nM. A final concentration of 300 nM for each primer is effective for most applications.Some primer set designs may require asymmetric primer concentrations.Always include a negative or no template control to evaluate the specificity of a given primer set and amplification protocol. Sequence specificity of the PCR should be confirmed by a method other than generation of a single melt peak. This includes confirmation of PCR product size, diagnostic restriction endonuclease fragment pattern, or sequencing.Preparation of a reaction cocktail is recommended to reduce pipetting errors and obtain reproducible HRM results. Assemble the reaction cocktail with all required components, except template DNA, and dispense equal aliquots into each reaction tube. Add DNA template as a final step. A minimum of 3 technical replicates for each DNA sample is recommended. Include appropriate positive controls for each sequence variant.Guidelines for PCR amplification and HRM analysis continued:Use approximately 10,000 copies of template DNA. A suggested input quantity for human genomic DNA is 10 to 30 ng.After sealing each reaction, vortex gently to mix contents. Centrifuge briefly to collect components at the bottom of the reaction tube.PCR amplification can be carried out in a conventional or real-time thermal cycler. Monitoring the reaction in real-time allows one to access the quality and PCR performance of a sample before HRM. All samples should produce comparable Cqs and fluorescence signal. Samples with delayed Cq (>30) or aberrant fluorescence signal should be excluded from the HRM analysis. Optimal cycling conditions will depend on the properties of your primers. Hold assembled reactions on ice, protected from light, if not proceeding immediately to PCR.Reaction AssemblyComponent Volume for 20-µL rxn. Final ConcentrationAccuMelt HRM SuperMix (2X) 10.0 µL 1xForward primer Variable 100 – 500 nMReverse primer Variable 100 – 500 nMNuclease-free water VariableDNA Template 2-5 µL ~10,000 copiesFinal Volume (µL) 20 µLPCR Cycling Protocol2-Step Cycling Protocol 3-Step Cycling Protocol Initial Denaturation 95ºC, 5 min*95ºC, 5 min*PCR cycling (40 to 45 cycles)Denaturation 95ºC, 5 to 10 s 95ºC, 5 to 10 sAnnealing60ºC, 30 s†15 s at 55 to 65ºCExtension 10 to 30s at 70ºC†HRM analysis§Consult instructions for your instrument* Full activation of AccuStart Taq DNA polymerase occurs within 30s at 95ºC; however, optimal initial denaturation time is template dependent and will affect PCR efficiency and sensitivity. Amplification of genomic DNA or supercoiled DNA targets may require 5 to 10 min at 95ºC to fully denature the template.† If monitoring PCR in real-time, collect and analyze kinetic PCR data at the end of the extension step. Extension time is dependent upon amplicon length and minimal data collection time requirement for your qPCR instrument. Some primer sets may require a 3-step cycling protocol for optimal performance. Optimal annealing temperature and time may need to be empirically determined for any given primer set.§ High Resolution Melting Analysis should be carried out immediately following PCR amplification. Please consult the instructions for your HRM instrument for procedural details. If not proceeding immediately to HRM, store plates at +4C protected from light. Mix and centrifuge samples immediately before HRM.Quality ControlKit components are free of contaminating DNase and RNase. AccuMelt HRM SuperMix is functionally tested to amplify a single copy gene in human genomic DNA and resolve homozygote samples for class 4 (A>T) and class 3 (C>G) SNPs in a model test system.Limited Label LicensesUse of this product signifies the agreement of any purchaser or user of the product to the following terms:1.The product may be used solely in accordance with the protocols provided with the product and this manual and for use with components contained in the kitonly. QIAGEN Beverly, Inc. grants no license under any of its intellectual property to use or incorporate the enclosed components of this kit with any components not included within this kit except as described in the protocols provided with the product, this manual, and additional protocols available at . Some of these additional protocols have been provided by Quantabio product users. These protocols have not been thoroughly tested or optimized by QIAGEN Beverly, Inc.. QIAGEN Beverly, Inc. neither guarantees them nor warrants that they do not infringe the rights of third-parties.2.Other than expressly stated licenses, QIAGEN Beverly, Inc. makes no warranty that this kit and/or its use(s) do not infringe the rights of third-parties.3.This kit and its components are licensed for one-time use and may not be reused, refurbished, or resold.4.QIAGEN Beverly, Inc. specifically disclaims any other licenses, expressed or implied other than those expressly stated.5.The purchaser and user of the kit agree not to take or permit anyone else to take any steps that could lead to or facilitate any acts prohibited above. QIAGEN Beverly,Inc. may enforce the prohibitions of this Limited License Agreement in any Court, and shall recover all its investigative and Court costs, including attorney fees, in any action to enforce this Limited License Agreement or any of its intellectual property rights relating to the kit and/or its components.The use of this product is covered by at least one claim of U.S. Patent No. 7,687,247 owned by Life Technologies Corporation. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product, (b) its components, or (c) materials made by the employment of this product or its components to a third party or otherwise use this product or its components or materials made by the employment of this product or its components for Commercial Purposes. Commercial Purposes means any activity for which a party receives or is due to receive consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. The buyer cannot use this product, or its components or materials made using this product or its components for therapeutic, diagnostic or prophylactic purposes. Further information on purchasing licenses under the above patents may be obtained by contacting the Licensing Department, Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, CA 92008. Email: *************************©2018 QIAGEN Beverly Inc. 100 Cummings Center Suite 407J Beverly, MA 01915Quantabio brand products are manufactured by QIAGEN, Beverly Inc.Intended for molecular biology applications. This product is not intended for the diagnosis, prevention or treatment of a disease.AccuMelt and AccuStart are trademarks of QIAGEN Beverly, Inc. SYTO is a registered trademark of Life Technologies Corporation (Molecular Probes Labeling and Detection Technologies).。

5种大黄蒽醌类衍生物的同时测定及应用引言大黄蒽醌类衍生物（anthraquinone derivative）是一类重要的天然产物。

它们被广泛应用于医学、化学、环保等领域。

同时测定不同大黄蒽醌衍生物的含量和质量是极其重要的，因为它们的化学结构和性质之间存在很大的差异。

本文将介绍5种大黄蒽醌类衍生物的同时测定及应用。

1. 大黄蒽醌大黄蒽醌（anthraquinone）是最简单的大黄蒽醌类衍生物。

它是一种有机化合物，化学式为C14H8O2。

大黄蒽醌广泛存在于花草、木材、棉麻、柑橘皮等天然产物中，并且在工业领域中也得到了广泛应用。

同时测定大黄蒽醌的含量非常重要，因为它有丰富的生物活性。

大黄蒽醌可以抑制某些致癌物质，缓解疼痛，以及对某些类胰岛素药物具有增强作用。

2. 芒果黄芒果黄（mangostin）是一种天然的大黄蒽醌衍生物，化学式为C24H26O6。

它广泛存在于热带地区的芒果果实中，并且在药物和化妆品领域中得到了广泛应用。

同时测定芒果黄的含量非常重要，因为它具有抗氧化、抗菌、抗炎等多种生物活性。

研究表明，芒果黄可以降低胆固醇、预防心血管疾病和糖尿病等慢性疾病的发生。

3. 大黄蒽醌-2-磺酸钠大黄蒽醌-2-磺酸钠（sodium anthraquinone-2-sulfonate）是一种合成的大黄蒽醌类衍生物，化学式为C14H8Na2O6S。

它是一种灰白色至白色粉末，可溶于水。

同时测定大黄蒽醌-2-磺酸钠的含量非常重要，因为它具有多种应用。

大黄蒽醌-2-磺酸钠被广泛应用于染料、颜料、纸张、医药等工业领域中。

4. 大黄蒽醌-2-甲醚磺酸钠大黄蒽醌-2-甲醚磺酸钠（sodium anthraquinone-2-methoxysulfonate）是一种合成的大黄蒽醌类衍生物，化学式为C15H11NaO6S。

它是一种灰白色至白色粉末，可溶于水。

同时测定大黄蒽醌-2-甲醚磺酸钠的含量非常重要，因为它被广泛应用于染料、颜料、纸张、医药等工业领域中。