Fast protein liquid chromatography

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…

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…

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…

Observe Ligand-Protein Interaction using Gel Filtration Chromatography (Phenol Red Binding to Bovine Serum Albumin) Introduction: Serum albumin is often known as blood albumin, and is the most abundant protein in humans and other animal plasma. Serum albumin is also known as a carrier protein by non-specifically binding several biomolecules. It is essential for maintaining the osmotic pressure that is needed for body fluids distribution between body tissues and intravascular compartments.…

This methodology varies from absorbtion, in which a liquid (the absorbate) pervades or is absorbed by a porous solid. Adsorption is a surface-based procedure while absorption includes the entire volume of the material. The term sorption envelops both methodologies, while desorption is the converse of it. Adsorption is the deposition of atoms or molecules on the surface of a substance. This creates a layer of the adsorbate (the molecules or atoms that are being collected) on the adsorbent…

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…

“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…

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…

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…

Chromatography- This is a technique that is most commonly used to separate a solution into its different components, an extraction solvent is used to do this, the solution is added to the chromatography paper, and then as the extraction solvent is absorbed by the paper the different compounds with different weights. How does it work- Chromatography is used to separate a mixture into its components. It contains two key phases, a stationary phase that involves a sold or a liquid supported in a…