Mixtures Are a Combination 3.1 of Two Or More Substances MIXTURES
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Protocols and Tips in Protein Purification
Department of Molecular Biology & Biotechnology Protocols and tips in protein purification or How to purify protein in one day Second edition 2018 2 Contents I. Introduction 7 II. General sequence of protein purification procedures 9 Preparation of equipment and reagents 9 Preparation and use of stock solutions 10 Chromatography system 11 Preparation of chromatographic columns 13 Preparation of crude extract (cell free extract or soluble proteins fraction) 17 Pre chromatographic steps 18 Chromatographic steps 18 Sequence of operations during IEC and HIC 18 Ion exchange chromatography (IEC) 19 Hydrophobic interaction chromatography (HIC) 21 Gel filtration (SEC) 22 Affinity chromatography 24 Purification of His-tagged proteins 25 Purification of GST-tagged proteins 26 Purification of MBP-tagged proteins 26 Low affinity chromatography 26 III. “Common sense” strategy in protein purification 27 General principles and tips in “common sense” strategy 27 Algorithm for development of purification protocol for soluble over expressed protein 29 Brief scheme of purification of soluble protein 36 Timing for refined purification protocol of soluble over -expressed protein 37 DNA-binding proteins 38 IV. Protocols 41 1. Preparation of the stock solutions 41 2. Quick and effective cell disruption and preparation of the cell free extract 42 3. Protamin sulphate (PS) treatment 43 4. Analytical ammonium sulphate cut (AM cut) 43 5. Preparative ammonium sulphate cut 43 6. Precipitation of proteins by ammonium sulphate 44 7. Recovery of protein from the ammonium sulphate precipitate 44 8. Analysis of solubility of expression 45 9. Analysis of expression for low expressed His tagged protein 46 10. Bio-Rad protein assay Sveta’s easy protocol 47 11. -
Water Specialist for the Oil & Gas Sector Ovivowater.Com
Water Specialist for the Oil & Gas Sector creating value in water through innovation, creativity and expertise ovivowater.com © 2013 GLV Inc. All rights reserved. Ovivo - Industrial Markets Ovivo: A market Leader As society and the global economy demand more and more from water, there is a growing requirement for ever more applications to manage clean water, to create specialist process waters, to treat wastewater, to extract energy from wastewater and to champion the reuse of water. Ovivo - creating value in water through innovation, creativity and Many of the best known, most respected and reliable expertise in clean water, process brand names in the water and wastewater industries are part of Ovivo’s heritage. The combined strengths water, wastewater treatment, waste- of our brands and talents, including Brackett Green, to-energy and water reuse markets Caird & Rayner Clark, Christ Water Technology and across 5 continents. Eimco Water Technologies create one of the most comprehensive bank of technologies in the sector, one of the most impressive list of references, and practical application knowledge. Ovivo aims to become the water partner of choice for clients in the public and private sectors and the leading source of water expertise for engineers and consultants across the globe. Get in touch with some of the best brains in the business. For further information, visit ovivowater.com Ovivo - bringing water to life Copyright © 2013 GLV Inc. All rights reserved. Water Specialist for the Oil & Gas Sector Our Role in your Industry Oil and gas are precious resources. As an Ovivo develops and implements innovative integral element of oil and gas production, water solutions to meet the challenges of treating water and management is especially important. -
Phase Transitions in Multicomponent Systems
Physics 127b: Statistical Mechanics Phase Transitions in Multicomponent Systems The Gibbs Phase Rule Consider a system with n components (different types of molecules) with r phases in equilibrium. The state of each phase is defined by P,T and then (n − 1) concentration variables in each phase. The phase equilibrium at given P,T is defined by the equality of n chemical potentials between the r phases. Thus there are n(r − 1) constraints on (n − 1)r + 2 variables. This gives the Gibbs phase rule for the number of degrees of freedom f f = 2 + n − r A Simple Model of a Binary Mixture Consider a condensed phase (liquid or solid). As an estimate of the coordination number (number of nearest neighbors) think of a cubic arrangement in d dimensions giving a coordination number 2d. Suppose there are a total of N molecules, with fraction xB of type B and xA = 1 − xB of type A. In the mixture we assume a completely random arrangement of A and B. We just consider “bond” contributions to the internal energy U, given by εAA for A − A nearest neighbors, εBB for B − B nearest neighbors, and εAB for A − B nearest neighbors. We neglect other contributions to the internal energy (or suppose them unchanged between phases, etc.). Simple counting gives the internal energy of the mixture 2 2 U = Nd(xAεAA + 2xAxBεAB + xBεBB) = Nd{εAA(1 − xB) + εBBxB + [εAB − (εAA + εBB)/2]2xB(1 − xB)} The first two terms in the second expression are just the internal energy of the unmixed A and B, and so the second term, depending on εmix = εAB − (εAA + εBB)/2 can be though of as the energy of mixing. -
Laboratory Filtration Product Guide
Laboratory filtration Product guide gelifesciences.com 1 Welcome to Whatman filtration by GE Healthcare Life Sciences Our reputation, based on a solid foundation of expertise, enables us to support how healthcare is researched and delivered. In laboratories across the globe, the Whatman™ name is synonymous with quality, reliability, and ease of use. Our instinct for simplification accelerates the rate of discovery, reduces costs and saves time. Our products have a reputation for working right the first time – every time, which is why they are specified for the most exacting applications across a wide range of industries for people around the globe. Basic analytical testing In the vast and disparate world of analytical chemistry, Whatman products are used for basic laboratory processes that range from simple clarification to solvent extraction. Products range from filter papers, thimbles and Benchkote™ benchtop protectors, to membrane filters and phase separator papers. Food and beverage Our filter papers are used to prepare food samples prior to a wide range of analyses. Our syringe filters prevent fatty or particulate laden samples from damaging valuable equipment. Our membranes are used to test for harmful bacteria. Pharmaceutical Whatman products enable pharmaceutical companies to increase productivity. Mini-UniPrep™ syringeless filters and vials reduce HPLC sample preparation time and consumables usage, and track-etched and Anopore™ membranes are also vital to extruding liposomes for encasing and targeting drugs. Environmental monitoring Whatman products are cited in EPA, ASTM and ISO protocols for environmental monitoring. Whether it is detecting suspended solids in water, measuring air for dangerous particulates, or supporting asbestos analysis to maintain healthy spaces there is a Whatman filter that is central to the test. -
Cellulose Nanopapers As Tight Aqueous Ultra- Filtration Membranes
Cellulose nanopapers as tight aqueous ultra- filtration membranes Andreas Mautner1*, Koon-Yang Lee2, Tekla Tammelin3, Aji P. Mathew4, Alisyn J. Nedoma5, Kang Li5 and Alexander Bismarck1,6,* 1 Polymer & Composite Engineering (PaCE) Group, Department of Chemical Engineering, Imperial College London, South Kensington Campus, SW7 2AZ London, United Kingdom 2 Department of Chemical Engineering, University College London, Torrington Place, WC1E 7JE London, United Kingdom 3 . VTT Technical Research Centre of Finland, Biologinkuja 7, FL-02044 Espoo, Finland 4 Division of Materials Science, Department of Engineering Sciences and Mathematics, Luleå University of Technology, 97187 Luleå, Sweden 5 Department of Chemical Engineering, Imperial College London, South Kensington Campus, SW7 2AZ London United Kingdom 6 Institute for Materials Chemistry & Research, Polymer & Composite Engineering (PaCE) Group, Faculty of Chemistry, University of Vienna, Währingerstr. 42, A-1090 Vienna, Austria * Corresponding authors: e-mail: [email protected], [email protected] Abstract: Recently, we have demonstrated the use of wood-derived nanocellulose papers, herein termed nanopapers, for organic solvent nanofiltration applications. In this study, we extend the use of these nanopapers to tight ultrafiltration (UF) membranes. The feasibility of such nanopaper-based UF membranes intended for use in water purification is shown. Four types of nanocelluloses, namely bacterial cellulose, wood-derived nanocellulose, TEMPO-oxidized cellulose nanofibrils and cellulose nanocrystals, were used as raw materials for the production of these nanopaper-based membranes. The resulting nanopapers exhibit a transmembrane permeance in the range of commercially available tight UF membranes with molecular weight cut-offs ranging from 6 to 25 kDa, which depends on the type of nanocellulose used. -
Grade 6 Science Mechanical Mixtures Suspensions
Grade 6 Science Week of November 16 – November 20 Heterogeneous Mixtures Mechanical Mixtures Mechanical mixtures have two or more particle types that are not mixed evenly and can be seen as different kinds of matter in the mixture. Obvious examples of mechanical mixtures are chocolate chip cookies, granola and pepperoni pizza. Less obvious examples might be beach sand (various minerals, shells, bacteria, plankton, seaweed and much more) or concrete (sand gravel, cement, water). Mechanical mixtures are all around you all the time. Can you identify any more right now? Suspensions Suspensions are mixtures that have solid or liquid particles scattered around in a liquid or gas. Common examples of suspensions are raw milk, salad dressing, fresh squeezed orange juice and muddy water. If left undisturbed the solids or liquids that are in the suspension may settle out and form layers. You may have seen this layering in salad dressing that you need to shake up before using them. After a rain fall the more dense particles in a mud puddle may settle to the bottom. Milk that is fresh from the cow will naturally separate with the cream rising to the top. Homogenization breaks up the fat molecules of the cream into particles small enough to stay suspended and this stable mixture is now a colloid. We will look at colloids next. Solution, Suspension, and Colloid: https://youtu.be/XEAiLm2zuvc Colloids Colloids: https://youtu.be/MPortFIqgbo Colloids are two phase mixtures. Having two phases means colloids have particles of a solid, liquid or gas dispersed in a continuous phase of another solid, liquid, or gas. -
Introduction to Phase Diagrams*
ASM Handbook, Volume 3, Alloy Phase Diagrams Copyright # 2016 ASM InternationalW H. Okamoto, M.E. Schlesinger and E.M. Mueller, editors All rights reserved asminternational.org Introduction to Phase Diagrams* IN MATERIALS SCIENCE, a phase is a a system with varying composition of two com- Nevertheless, phase diagrams are instrumental physically homogeneous state of matter with a ponents. While other extensive and intensive in predicting phase transformations and their given chemical composition and arrangement properties influence the phase structure, materi- resulting microstructures. True equilibrium is, of atoms. The simplest examples are the three als scientists typically hold these properties con- of course, rarely attained by metals and alloys states of matter (solid, liquid, or gas) of a pure stant for practical ease of use and interpretation. in the course of ordinary manufacture and appli- element. The solid, liquid, and gas states of a Phase diagrams are usually constructed with a cation. Rates of heating and cooling are usually pure element obviously have the same chemical constant pressure of one atmosphere. too fast, times of heat treatment too short, and composition, but each phase is obviously distinct Phase diagrams are useful graphical representa- phase changes too sluggish for the ultimate equi- physically due to differences in the bonding and tions that show the phases in equilibrium present librium state to be reached. However, any change arrangement of atoms. in the system at various specified compositions, that does occur must constitute an adjustment Some pure elements (such as iron and tita- temperatures, and pressures. It should be recog- toward equilibrium. Hence, the direction of nium) are also allotropic, which means that the nized that phase diagrams represent equilibrium change can be ascertained from the phase dia- crystal structure of the solid phase changes with conditions for an alloy, which means that very gram, and a wealth of experience is available to temperature and pressure. -
Gp-Cpc-01 Units – Composition – Basic Ideas
GP-CPC-01 UNITS – BASIC IDEAS – COMPOSITION 11-06-2020 Prof.G.Prabhakar Chem Engg, SVU GP-CPC-01 UNITS – CONVERSION (1) ➢ A two term system is followed. A base unit is chosen and the number of base units that represent the quantity is added ahead of the base unit. Number Base unit Eg : 2 kg, 4 meters , 60 seconds ➢ Manipulations Possible : • If the nature & base unit are the same, direct addition / subtraction is permitted 2 m + 4 m = 6m ; 5 kg – 2.5 kg = 2.5 kg • If the nature is the same but the base unit is different , say, 1 m + 10 c m both m and the cm are length units but do not represent identical quantity, Equivalence considered 2 options are available. 1 m is equivalent to 100 cm So, 100 cm + 10 cm = 110 cm 0.01 m is equivalent to 1 cm 1 m + 10 (0.01) m = 1. 1 m • If the nature of the quantity is different, addition / subtraction is NOT possible. Factors used to check equivalence are known as Conversion Factors. GP-CPC-01 UNITS – CONVERSION (2) • For multiplication / division, there are no such restrictions. They give rise to a set called derived units Even if there is divergence in the nature, multiplication / division can be carried out. Eg : Velocity ( length divided by time ) Mass flow rate (Mass divided by time) Mass Flux ( Mass divided by area (Length 2) – time). Force (Mass * Acceleration = Mass * Length / time 2) In derived units, each unit is to be individually converted to suit the requirement Density = 500 kg / m3 . -
Partition Coefficients in Mixed Surfactant Systems
Partition coefficients in mixed surfactant systems Application of multicomponent surfactant solutions in separation processes Vom Promotionsausschuss der Technischen Universität Hamburg-Harburg zur Erlangung des akademischen Grades Doktor-Ingenieur genehmigte Dissertation von Tanja Mehling aus Lohr am Main 2013 Gutachter 1. Gutachterin: Prof. Dr.-Ing. Irina Smirnova 2. Gutachterin: Prof. Dr. Gabriele Sadowski Prüfungsausschussvorsitzender Prof. Dr. Raimund Horn Tag der mündlichen Prüfung 20. Dezember 2013 ISBN 978-3-86247-433-2 URN urn:nbn:de:gbv:830-tubdok-12592 Danksagung Diese Arbeit entstand im Rahmen meiner Tätigkeit als wissenschaftliche Mitarbeiterin am Institut für Thermische Verfahrenstechnik an der TU Hamburg-Harburg. Diese Zeit wird mir immer in guter Erinnerung bleiben. Deshalb möchte ich ganz besonders Frau Professor Dr. Irina Smirnova für die unermüdliche Unterstützung danken. Vielen Dank für das entgegengebrachte Vertrauen, die stets offene Tür, die gute Atmosphäre und die angenehme Zusammenarbeit in Erlangen und in Hamburg. Frau Professor Dr. Gabriele Sadowski danke ich für das Interesse an der Arbeit und die Begutachtung der Dissertation, Herrn Professor Horn für die freundliche Übernahme des Prüfungsvorsitzes. Weiterhin geht mein Dank an das Nestlé Research Center, Lausanne, im Besonderen an Herrn Dr. Ulrich Bobe für die ausgezeichnete Zusammenarbeit und der Bereitstellung von LPC. Den Studenten, die im Rahmen ihrer Abschlussarbeit einen wertvollen Beitrag zu dieser Arbeit geleistet haben, möchte ich herzlichst danken. Für den außergewöhnlichen Einsatz und die angenehme Zusammenarbeit bedanke ich mich besonders bei Linda Kloß, Annette Zewuhn, Dierk Claus, Pierre Bräuer, Heike Mushardt, Zaineb Doggaz und Vanya Omaynikova. Für die freundliche Arbeitsatmosphäre, erfrischenden Kaffeepausen und hilfreichen Gespräche am Institut danke ich meinen Kollegen Carlos, Carsten, Christian, Mohammad, Krishan, Pavel, Raman, René und Sucre. -
Liquid / Solids Separation in Wastewater Treatment & Biosolids Dewatering
LIQUID / SOLIDS SEPARATION IN WASTEWATER TREATMENT & BIOSOLIDS DEWATERING Chemical Products Lab Testing Plant Trials LIQUID / SOLIDS SEPARATION APPLICATIONS Influent Water Clarification Process Water Recycling Primary Wastewater Clarification Secondary Clarification Sludge Thickening Sludge Dewatering LIQUID / SOLIDS SEPARATION UNIT OPERATIONS Clarifiers (Many Types) WATER Filters (Many Types) OR WASTE Dissolved Air Flotation Units WATER Induced Air/Gas Flotation Units Belt Presses Centrifuges SLUDGE Screw Presses DEWATERING Plate and Frame Presses Vacuum Filters (Rotary & Horizontal) LIQUID / SOLIDS SEPARATION PRODUCT TYPES Coagulants (+) Low Mol Wt Organic Inorganic Blended Flocculants (+ , ---, 0 ) High Mol Wt Dry Emulsion Solution OilOil----FreeFree Flocculants COAGULANTS AND FLOCCULANTS Act on Insoluble Particles in Water Oils, Grease, Blood, Insoluble Organics, Clay, Silicates, Metal Oxides/Hydroxides Dirt, Dust, Rust & Metal Filings Can Act on Charged Organic Compounds Anionic Surfactants, Soaps & Dispersants Do Not Act on Most Dissolved Solids Salts, Acids, Nonionic Surfactants, Ammonia or Soluble Organic Compounds such as Sugar, Alcohols, etc. SUSPENSION CHEMISTRY THE KEY TO EFFECTIVE LIQUID / SOLIDS SEPARATION SUSPENDED SOLIDS VARIABLES Surface Charge MOST Charge Density Particle Size IMPORTANCE Composition Particle Density Particle Shape LEAST MICROSCOPIC FORCES ELECTROSTATIC BROWNIAN VAN DER WAALS GRAVITY Colloidal Particle in Water +++ +++ +++ +++ +++ +++ +++ +++ +++ +++ Almost all Particles +++ -
Ways to Physically Separate a Mixture There Are 2 Types of Mixtures
Mixtures 12 Mixtures Homogenous Compare Heterogeneous compounds and Suspension Mixtures. Colloid Solution Differentiate Solute/Solvent between solutions, suspensions, and colloids Comparing Mixtures and Compounds Mixtures Compounds Made of 2 or more substances Made of 2 or more substances physically combined chemically combined Substances keep their own properties Substances lose their own properties Can be separated by physical means Can be separated only by chemical means Have no definite chemical composition Have a definite chemical composition What method is used to separate mixtures Ways to physically separate a Mixture based on boiling • Distillation – uses boiling point point? • Magnet – uses magnetism What method is used • Centrifuge – uses density to separate mixtures based on density? • Filtering – separates large particles from smaller ones What method is used There are 2 types of mixtures: to separate mixtures (1) Heterogeneous Mixtures: the parts mixed together can still be based on particle size? distinguished from one another...NOT uniform in composition Give examples of a heterogeneous Examples: chicken soup, fruit salad, dirt, sand in water mixture Mixtures 12 (2) Homogenous Mixtures: the parts mixed together cannot be distinguished from one another...completely uniform in composition. Give examples of a homogenous Examples: Air, Kool-aid, Brass, salt water, milk mixture Differentiate Types of Homogenous mixtures between a homogenous 1. Suspensions mixture and a i.e. chocolate milk, muddy water, Italian dressing heterogeneous mixture. They are cloudy (usually a liquid mixed with small solid particles) Identify an example of a suspension. Needs to be shaken or stirred to keep the solids from Will the solid settling particles settle in a suspension? The solids can be filtered out 2. -
Solving Mixture and Solution Verbal Problems
SOLVING SOLUTION AND MIXTURE VERBAL PROBLEMS This type of problem involves mixing two different solutions of a certain ingredient to get a desired concentration of the ingredient. Before we can solve problems that involve concentrations, we must review certain concepts about percents. If you need to do this, go to the brush-up materials for solving percent problems on the Dolciani website. 1. Solution Problems Basic Equation: amount of solution concentration of substance = amount of substance Example: 40 ounces (amount of solution) of a 25% solution of acid (concentration) contains 25(40) = 10 ounces of acid Usual equation to solve for the variable: Amount of substance in solution 1 + Amount of substance in solution 2 = Amount of substance in solution 2. Mixture Problems Basic Equation: unit price # units = cost (or value) Example: 5 pounds of apples (# units) that sell for $1.20 per pound (unit price) costs 5(1.20) = $6 Using equation to solve for the variable: Cost of ingredient 1 + Cost of ingredient 2 = Cost of mixture Now let’s look at an actual mixture problem. It is easiest when solving a mixture problem to make a table to get the information organized. No matter the story line of the problem, the table can be used and labelled as necessary. Let’s look at a few examples. Example 1: Fatima’s chemistry lab stocks an 8% acid solution and a 20% acid solution. How many ounces of each must she combine to produce 60 ounces of a mixture that is 10% acid? Solution: We want to find how much of each solution must be in the mixture.