Fast protein liquid chromatography

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    I conducted the purification of the protein under the guidance of Dr. Raquel Ruiz and involved expression tests at different concentrations of the inducing agent IPTG, large-scale growth of BL21-DE3 E. coli cells that over-express hS100A7, utilizing fast protein liquid chromatography (FPLC) with immobilized metal ion affinity and size exclusion columns, and ultimately using the purified protein product in setting up crystallization screens. I was unable to obtain any crystals of hS100A7 due to the usage of the protease inhibitor cocktail AEBSF after cell lysis, which is known to interfere with protein crystallization. The next steps after storage of the frozen protein are to remove any endotoxins in the protein solution in order to be used in in vivo drug…

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    ten-minute interval. These samples were then sonicated using a Branson 8570 sonicating bath. The samples sat in the sonicating bath for a five-minute interval. The acetaminophen samples were then transferred to centrifuge tubes (roughly 12mL of solution) and set into a VWR Clinical 50 Centrifuge at 4000rpm for five minutes. After five minutes, half of the solution, with none of the solid from the bottom of the centrifuge tube, was pipetted into another centrifuge tube. These solutions then had…

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    Chromatography is a process used to separate molecules on the basis of chemical or physical properties, such as molecular mass, charge, or hydrophobicity. (Ninfa 121) There are many types of chromatography processes that can be used in different experiments and scenarios. These chromatography processes on include Gel filtration, ion exchange, and high pressure liquid chromatography. However, in this laboratory session the process of affinity chromatography is utilized to purify a His-tagged…

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    3. Results and discussion 3.1 Microstructural characterization The Roman spectrum of Cr-DLC coating was shown in Fig.1. The overlapped D peak and G peaks located at 1363cm-1 and 1564cm-1 were obtained after fitted with Gaussian curves, respectively. It’s well known that the content of the graphite crystal was closely related to the value of ID/IG [22]. And the ratio of intensity between D peak and G peak (ID/IG) of Cr-DLC coating could be calculated by the fitted curve. The value of ID/IG was…

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    “DEVELOPMENT AND VALIDATION OF STABILITY INDICATING METHOD FOR SIMULTANEOUS ESTIMATION OF CEFTRIAXONE AND TAZOBACTAM INJECTION USING RP-UPLC METHOD” 2.1 OBJECTIVE To develop and Validate Stability Indicating Method for simultaneous estimation of Ceftriaxone and Tazobactam in their combined dosage form by RP-UPLC method in various stress condition like, alkaline, acidic, oxidative, thermal and photo degradation. 2.4 METHODOLOGY 2.4.1 Chromatographic Condition Column Separation was achieved by…

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    Liquid Chromatography Lab

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    High performance liquid chromatography (HPLC) is a separation technique utilizing differences in distribution of compounds in two phases; called the stationary phase and the mobile phase. The stationary phase designates a thin layer created on the surface of fine particles and the mobile phase designates the liquid flowing over the particles. Under a certain dynamic condition, each component in a sample has a different distribution equilibrium depending upon the solubility in the phases and or…

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    Uplc Water Chromatography

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    The chromatography was performed on an ACQUITY TM UPLC system (Waters Corp., Milford, MA, USA). The UPLC system included quaternary solvent manager, a binary pump, degasser, autosampler with an injection loop of 10 μL and a column heater-cooler. The separation was performed on Acquity UPLC BEH TM C18 column (50 × 2.1 mm, i.d., 1.7 μm, Waters, USA) maintained at 40 °C. a mobile phase of acetonitrile containing 0.1% formic acid and aqueous 5 mM ammonium acetate buffer containing 0.1% formic acid…

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    Keywords: Aralia continetalis Kitagawa; Kaurenoic acid; Phase I; Pharmacokinetics; Ultra-performance liquid chromatography-tandem mass spectrometry Abbreviations: AIC, akaike information criteria; AUC0-∞, the area under the concentration–time curve from zero to infinity; CL/F, the total body clearance for oral administration; Cmax, the maximum plasma concentration; CV, coefficient of variation; HPLC, high-performance liquid chromatography; ka, the absorption rate constant; IS, internal…

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    Introduction: Centrifugation is a process used to separate or concentrate materials suspended in a liquid medium. It is a method used to separate molecules under the centrifugation field based on their sedimentation rate with the help of centrifugal force. It is also used to measure the physical properties of a molecule like molecular weight, density, shape, etc. If it is used for the separation of one type of materials from others is termed as preparative centrifugation and when used for…

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    Chromatography is a method of separation in which the components of a substance are separated and distributed between two phases, which are the mobile phases and stationary phase (1). There are multiple types of chromatograph, which includes paper chromatography, column chromatograph, retention chromatography, gas chromatography, liquid chromatography etc. Each technique has its own strengths and weaknesses, but all are useful in getting separations of different components (8). Since many…

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