Marmara Üniversitesi Fen-Edebiyat Fakültesi Kimya Bölümü / Biyokimya Anabilim Dalı PURIFICATION AND CHARACTERIZATION OF PROTEINS
|
|
- Nelson Golden
- 8 years ago
- Views:
Transcription
1 EXPERIMENT VI PURIFICATION AND CHARACTERIZATION OF PROTEINS I- Protein isolation and dialysis In order to investigate its structure and properties a protein must be obtained in pure form. Since proteins are easily denaturated and found as a complex mixture in biological materials (such as body fluids and tissue extracts), their purification is quite difficult. Source for obtaining proteins are usually tissues and cells. First step in any purification process is destroying (or lysing) the cell and solubilize its protein content in lysis buffer. This solution is called as crude extract and it contains both targeted protein and all the other molecules in the cell that soluble in chosen lysis buffer. Later, crude protein extract fractioned according their different properties such as solubility, size, electrical charge, and polarity. Various separation techniques could be use in that purpose; differential centrifuge, salting out, precipitation with cold acetone or alcohol, ultrafiltration, and various chromatographic methods. A sample methodology for purification of an enzyme from crude extract is given below: a- Obtaining crude extract using a lysis buffer b- Salting out using ammonium sulfate (solubility in salt solutions) c- Ion exchange chromatography (electrical charge difference) d- Size exclusion (molecular sieve) chromatography e- Affinity chromatography (using a specifically binding ligand) Proteins become more and more pure in each purification step although they lost activity. When working with enzyme ph and temperature of the solutions should be controlled carefully. Process Step Fraction volume (ml) Total Protein (mg) Activity (Unit) Crude extract Salting out Spesific activity(unit/mg) Ion exchange Size exclusion Affinity Biyokimya Laboratuvarı I Güz 0
2 Ion Exchange Chormatography Marmara Üniversitesi Biyokimya Laboratuvarı I Güz 1
3 Size Exclusion (Molecular Sieve) Chromatography Biyokimya Laboratuvarı I Güz 2
4 Affinity Chromatography Marmara Üniversitesi Biyokimya Laboratuvarı I Güz 3
5 Salt ions that remains in the solution after the salting out interferes with the following experiments and therefore should be removed. Gel filtration, ultracentrifuge and dialysis could be used for that purpose. Dialysis is a separation technique that facilitates the removal of small, unwanted compounds from macromolecules in solution by selective and passive diffusion through a semi-permeable membrane. Membranes used in dialysis are made of cellulose and have definite pore size (usually Da). Salts at low concentration usually used as dialysis buffer. According to the rule of diffusion salts in the protein extract move to the outside of the membrane until concentration is equal both outside and inside of the dialysis tubing. However, since they are much bigger than salt molecules, proteins stay inside the dialysis sack. Equilibrium is reached approximately in 4-6 hours. At this point, dialysis buffer replaced with a fresh one. Dialysis usually complete in hours. CHEMICALS Potassium dihydrogen phosphate, disodium hydrogen phosphate, ammonium sulfate, urea, thiourea, trishydroxymethyl aminomethane (Tris), sodium dodecyl sulfate (SDS), dialysis tubings. METHOD In this experiments proteins extracted from 3 different sources; liver, egg yolk and egg whites. Biyokimya Laboratuvarı I Güz 4
6 Lysis Buffer: 9 M urea, 0.04 M Tris and 0.1 SDS% g Tris and g urea dissolved in a 100 ml volumetric flask using ph 7.4 Sørensen buffer. Homogenization Wash the liver, and get it into small pieces with lysis buffer in laboratory blender. Further homogenize it using a homogenizer. Add 1 ml 10% SDS to the homogenate and centrifuge. Supernatant decanted in to a beaker. Take egg yolk and egg white in a beaker and add lysis buffer and small amount of 10% SDS. Carefully mix it, centrifuge and collect supernatant into a beaker. Precipitation Salting out: Take 10 ml of supernatant into a plastic centrifuge tube and using saturation table add required amount of ammonium sulfate. Centrifuge and discard the supernatant. Precipitate will be used in further steps. Precipitation with acetone: Take 4 ml of supernatant into a plastic centrifuge tube and add 8 ml of cold acetone. Mix carefully, centrifuge and discard the supernatant. Precipitate will be used in further steps. Solubilization Add minumun amount of ph7.4 Sørensen phosphate buffer to the precipitate and carefully mix to solve proteins. Biyokimya Laboratuvarı I Güz 5
7 Dialysis After salting out step, solubilized proteins dialyzed against distilled water at cold for 2 days, distilled water refreshed every 4 hours. Protein solutions store at -80 C. Question 1- How proteins can be isolated from natural sources? Examine the scientific article and write down what kind of methods used in that experiment. Biyokimya Laboratuvarı I Güz 6
6 Characterization of Casein and Bovine Serum Albumin
6 Characterization of Casein and Bovine Serum Albumin (BSA) Objectives: A) To separate a mixture of casein and bovine serum albumin B) to characterize these proteins based on their solubilities as a function
More informationHiPer Ion Exchange Chromatography Teaching Kit
HiPer Ion Exchange Chromatography Teaching Kit Product Code: HTC001 Number of experiments that can be performed: 5 Duration of Experiment: Protocol: 5-6 hours Storage Instructions: The kit is stable for
More informationTECHNICAL BULLETIN. HIS-Select Nickel Affinity Gel. Catalog Number P6611 Storage Temperature 2 8 C
HIS-Select Nickel Affinity Gel Catalog Number P6611 Storage Temperature 2 8 C TECHNICAL BULLETIN Product Description HIS-Select Nickel Affinity Gel is an immobilized metalion affinity chromatography (IMAC)
More informationDNA SPOOLING 1 ISOLATION OF DNA FROM ONION
DNA SPOOLING 1 ISOLATION OF DNA FROM ONION INTRODUCTION This laboratory protocol will demonstrate several basic steps required for isolation of chromosomal DNA from cells. To extract the chromosomal DNA,
More informationGenomic DNA Extraction Kit INSTRUCTION MANUAL
Genomic DNA Extraction Kit INSTRUCTION MANUAL Table of Contents Introduction 3 Kit Components 3 Storage Conditions 4 Recommended Equipment and Reagents 4 Introduction to the Protocol 4 General Overview
More informationISOLATION OF CAFFEINE FROM TEA
ISLATIN F CAFFEINE FRM TEA Introduction In this experiment, caffeine is isolated from tealeaves. The chief problem with the isolation is that caffeine does not exist alone in the tealeaves, but other natural
More informationGuide to Reverse Phase SpinColumns Chromatography for Sample Prep
Guide to Reverse Phase SpinColumns Chromatography for Sample Prep www.harvardapparatus.com Contents Introduction...2-3 Modes of Separation...4-6 Spin Column Efficiency...7-8 Fast Protein Analysis...9 Specifications...10
More informationPlant Genomic DNA Extraction using CTAB
Plant Genomic DNA Extraction using CTAB Introduction The search for a more efficient means of extracting DNA of both higher quality and yield has lead to the development of a variety of protocols, however
More informationClassic Immunoprecipitation
292PR 01 G-Biosciences 1-800-628-7730 1-314-991-6034 technical@gbiosciences.com A Geno Technology, Inc. (USA) brand name Classic Immunoprecipitation Utilizes Protein A/G Agarose for Antibody Binding (Cat.
More informationHow To Make A Tri Reagent
TRI Reagent For processing tissues, cells cultured in monolayer or cell pellets Catalog Number T9424 Store at room temperature. TECHNICAL BULLETIN Product Description TRI Reagent is a quick and convenient
More informationExperiment #10: Liquids, Liquid Mixtures and Solutions
Experiment #10: Liquids, Liquid Mixtures and Solutions Objectives: This experiment is a broad survey of the physical properties of liquids. We will investigate solvent/solute mixtures. We will study and
More informationAffi-Prep Protein A Matrix Instruction Manual
Affi-Prep Protein A Matrix Instruction Manual Catalog Numbers 156-0005 156-0006 Bio-Rad Laboratories, 2000 Alfred Nobel Dr., Hercules, CA 94547 LIT-230 Rev B Table of Contents Section 1 Introduction...1
More informationBenchtop Mitochondria Isolation Protocol
Benchtop Mitochondria Isolation Protocol Note: Specific protocols are available for the following products: MS850 Mitochondria Isolation Kit for Rodent Tissue MS851 Mitochondria Isolation Kit for Rodent
More informationLAB 11 PLASMID DNA MINIPREP
LAB 11 PLASMID DNA MINIPREP STUDENT GUIDE GOAL The objective of this lab is to perform extraction of plasmid DNA and analyze the results. OBJECTIVES After completion, the student should be able to: 1.
More informationBiology 29 Cell Structure and Function Spring, 2009 Springer LABORATORY 2:CHLOROPLASTS AND PHOTOREDUCTION
Biology 29 Cell Structure and Function Spring, 2009 Springer LABORATORY 2:CHLOROPLASTS AND PHOTOREDUCTION In this laboratory we will purify chloroplasts from spinach by differential centrifugation, then
More informationPure-IP Western Blot Detection Kit
Product Manual Pure-IP Western Blot Detection Kit Catalog Number PRB-5002 20 blots FOR RESEARCH USE ONLY Not for use in diagnostic procedures Introduction The technique of immunoprecipitation (IP) is used
More informationPharmaceutical Biotechnology. Recombinant DNA technology Western blotting and SDS-PAGE
Pharmaceutical Biotechnology Recombinant DNA technology Western blotting and SDS-PAGE Recombinant DNA Technology Protein Synthesis Western Blot Western blots allow investigators to determine the molecular
More informationProtein extraction from Tissues and Cultured Cells using Bioruptor Standard & Plus
Protein extraction from Tissues and Cultured Cells using Bioruptor Standard & Plus Introduction Protein extraction from tissues and cultured cells is the first step for many biochemical and analytical
More informationCorning Spin-X UF Concentrator Selection and Use Guide
Corning Spin-X UF Concentrator Selection and Use Guide Contents Introduction.......................................................... 2 Choosing the Right Concentrator......................................
More informationChapter 3 Contd. Western blotting & SDS PAGE
Chapter 3 Contd. Western blotting & SDS PAGE Western Blot Western blots allow investigators to determine the molecular weight of a protein and to measure relative amounts of the protein present in different
More informationApplication Guide... 2
Protocol for GenomePlex Whole Genome Amplification from Formalin-Fixed Parrafin-Embedded (FFPE) tissue Application Guide... 2 I. Description... 2 II. Product Components... 2 III. Materials to be Supplied
More informationGenomic DNA Purification Student Laboratory Manual
Genomic DNA Purification Student Laboratory Manual I. Purpose...1 II. Introduction...1 III. DNA Purification from Bananas...2 IV. DNA Purification from Tissue Culture Cells...4 I. Purpose! This student
More informationBio-Gel P Polyacrylamide Gel Instruction Manual
Bio-Gel P Polyacrylamide Gel Instruction Manual Table of Contents Section 1 Introduction...1 Section 2 Technical Description...3 Section 3 Instructions for Use...6 3.1 Column Selection...6 3.2 Eluant Selection...6
More informationCatalase. ***You will be working with hot water, acids and bases in this laboratory*** ****Use Extreme Caution!!!****
AP BIOLOGY BIOCHEMISTRY ACTIVITY #9 NAME DATE HOUR CATALASE LAB INTRODUCTION Hydrogen peroxide (H 2 O 2 ) is a poisonous byproduct of metabolism that can damage cells if it is not removed. Catalase is
More informationAnn.wellhouse@TouchStoneScience.net 1. Enzyme Function
Ann.wellhouse@TouchStoneScience.net 1 Enzyme Function National Science Standards Science as Inquiry: Content Standard A: As a result of activities in grades 9-12, all students should develop: Abilities
More informationChapter 5, Lesson 3 Why Does Water Dissolve Salt?
Chapter 5, Lesson 3 Why Does Water Dissolve Salt? Key Concepts The polarity of water molecules enables water to dissolve many ionically bonded substances. Salt (sodium chloride) is made from positive sodium
More informationPurification of Plasmid DNA
Purification of Plasmid DNA Introduction: The growth of colonies on antibiotic medium provides phenotypic evidence that cells have been transformed. To confirm this at the genotypic level, plasmid DNA
More informationRiboZol RNA Extraction Reagents
RiboZol RNA Extraction Reagents Code Description Size N580-30ML-SAMPLE Ribozol TM RNA Extraction Reagent 30 ml N580-30ML Ribozol TM RNA Extraction Reagent 30 ml N580-100ML Ribozol TM RNA Extraction Reagent
More informationProtein purification methods, a practical approach
r i Protein purification methods, a practical approach 2008 AGI-Information Management Consultants May be used for personal purporses only or by libraries associated to dandelon.com network. I Edited by
More informationPCR and Sequencing Reaction Clean-Up Kit (Magnetic Bead System) 50 preps Product #60200
3430 Schmon Parkway Thorold, ON, Canada L2V 4Y6 Phone: 866-667-4362 (905) 227-8848 Fax: (905) 227-1061 Email: techsupport@norgenbiotek.com PCR and Sequencing Reaction Clean-Up Kit (Magnetic Bead System)
More informationReadyPrep Protein Extraction Kit (Cytoplasmic/Nuclear)
ReadyPrep Protein Extraction Kit (Cytoplasmic/Nuclear) Instruction Manual Catalog #163-2089 For technical service, call your local Bio-Rad office, or in the US, call 1-800-4BIORAD (1-800-424-6723) Bio-Rad
More informationUltraClean Soil DNA Isolation Kit
PAGE 1 UltraClean Soil DNA Isolation Kit Catalog # 12800-50 50 preps New improved PCR inhibitor removal solution (IRS) included Instruction Manual (New Alternative Protocol maximizes yields) Introduction
More informationExpression and Purification of Recombinant Protein in bacteria and Yeast. Presented By: Puspa pandey, Mohit sachdeva & Ming yu
Expression and Purification of Recombinant Protein in bacteria and Yeast Presented By: Puspa pandey, Mohit sachdeva & Ming yu DNA Vectors Molecular carriers which carry fragments of DNA into host cell.
More informationEXTRACTION OF DNA FROM CALF THYMUS CELLS Revised 2/1/96 Introduction
Revised 2/1/96 Introduction Cells may be classified into two primary types depending on whether they have a discrete nucleus (eukaryotic) or do not (prokaryotic). Prokaryotes include bacteria, such as
More informationReading instructions to partitioning in aqueous two-phase systems
Reading instructions to partitioning in aqueous two-phase systems Copy from Separation Processes in Biotechnology (ed. Asenjo) Aqueous two-phase separations, Albertsson, Johansson och Tjerneld OH-pictures
More informationPreparation of frequently used solutions
Preparation of frequently used solutions Content 1. Diluting Concentrated Acids (Last Login: 08/08/2009) 2. Indicators (Last Login: 27/07/2009) 3. Standard Buffer Solutions (Last Login: 27/07/2009) 4.
More informationSODIUM CARBOXYMETHYL CELLULOSE
SODIUM CARBOXYMETHYL CELLULOSE Prepared at the 28th JECFA (1984), published in FNP 31/2 (1984) and in FNP 52 (1992). Metals and arsenic specifications revised at the 55 th JECFA (2000). An ADI not specified
More informationHighPure Maxi Plasmid Kit
HighPure Maxi Plasmid Kit For purification of high pure plasmid DNA with high yields www.tiangen.com PP120109 HighPure Maxi Plasmid Kit Kit Contents Storage Cat.no. DP116 Contents RNaseA (100 mg/ml) Buffer
More informationFactors Affecting Enzyme Activity
INTRODUCTION Factors Affecting Enzyme Activity The chemical reactions occurring in living things are controlled by enzymes. An enzyme is a protein in the cell which lowers the activation energy of a catalyzed
More informationOSMOSIS AND DIALYSIS 2003 BY Wendy Weeks-Galindo with modifications by David A. Katz
OSMOSIS AND DIALYSIS 2003 BY Wendy Weeks-Galindo with modifications by David A. Katz OSMOSIS Osmosis is the reason that a fresh water fish placed in the ocean desiccates and dies. Osmosis is the reason
More informationSTABILITY: TRI REAGENT is stable at 25 C for at least two years from the date of purchase (3).
PRODUCT: TRI REAGENT - RNA / DNA / PROTEIN ISOLATION REAGENT May 2014 Cat. No: TR 118 Storage: Store at 4-25 C PRODUCT DESCRIPTION TRI REAGENT is a complete and ready-to-use reagent for the isolation of
More informationMethods of Grading S/N Style of grading Percentage Score 1 Attendance, class work and assignment 10 2 Test 20 3 Examination 70 Total 100
COURSE: MIB 303 Microbial Physiology and Metabolism (3 Units- Compulsory) Course Duration: Three hours per week for 15 weeks (45 hours). Lecturer: Jimoh, S.O. B.Sc., M.Sc, Ph.D Microbiology (ABU, Zaria)
More informationFIGURE 2.18. A. The phosphate end of the molecule is polar (charged) and hydrophilic (attracted to water).
PLASMA MEMBRANE 1. The plasma membrane is the outermost part of a cell. 2. The main component of the plasma membrane is phospholipids. FIGURE 2.18 A. The phosphate end of the molecule is polar (charged)
More informationProcess of Science: Using Diffusion and Osmosis
Process of Science: Using Diffusion and Osmosis OBJECTIVES: 1. To understand one way to approach the process of science through an investigation of diffusion and osmosis. 2. To explore how different molecules
More informationWestern Blotting. Prepare samples:
Western Blotting Sive Lab Protocol March 2007 Prepare samples: For zebrafish embryos: Option 1: Take live embryos and put into 1.5 ml tube with E3. Centrifuge gently for 1-2 minutes -yolk lipids will rise
More informationcatalase 2H 2 O 2 (l) ----> 2H 2 O (l) + O 2 (g)
ENZYME POST LAB QUIZ STUDY GUIDE Below are the answers to the post-lab (Data Analysis) questions. Make sure you UNDERSTAND all of these questions. The post-lab questions will, of course, be different,
More informationBasics of SPE Technology & Mechanisms. Pieter Grobler, Sigma-Aldrich RSA
Basics of SPE Technology & Mechanisms Pieter Grobler, Sigma-Aldrich RSA Agenda The Importance of Sample Prep Overview of SPE Technology SPE Strategies Understanding Retention Mechanisms Analytical Chromatography
More information--not necessarily a protein! (all proteins are polypeptides, but the converse is not true)
00Note Set 5b 1 PEPTIDE BONDS AND POLYPEPTIDES OLIGOPEPTIDE: --chain containing only a few amino acids (see tetrapaptide, Fig 5.9) POLYPEPTIDE CHAINS: --many amino acids joined together --not necessarily
More informationTOTAL PROTEIN FIBRINOGEN
UNIT: Proteins 16tproteins.wpd Task Determination of Total Protein, Albumin and Globulins Objectives Upon completion of this exercise, the student will be able to: 1. Explain the ratio of albumin and globulin
More informationMagExtractor -Genome-
Instruction manual MagExtractor-Genome-0810 F0981K MagExtractor -Genome- NPK-101 100 preparations Store at 4 C Contents [1] Introduction [2] Components [3] Materials required [4] Protocol 1. Purification
More informationHonors 227 Fall 2007 Laboratory with Ms. Clark. Enzymes, Reactions, Metabolism and Homeostasis
1 Name: Honors 227 Fall 2007 Laboratory with Ms. Clark Enzymes, Reactions, Metabolism and Homeostasis Background Enzymes, which are comprised of amino acids, are very important macromolecules found in
More informationMolecular Cell Biology. Prof. D. Karunagaran. Department of Biotechnology. Indian Institute of Technology Madras
Molecular Cell Biology Prof. D. Karunagaran Department of Biotechnology Indian Institute of Technology Madras Module 5 Methods in Cell Biology (Methods to Manipulate Protein, DNA and RNA and Methods to
More informationPerfectFOCUS. For Preparing Low Conductivity Samples for IEF/2D-Gel Electrophoresis. (Cat. # 786-124, 786-124T)
357PR G-Biosciences 1-800-628-7730 1-314-991-6034 technical@gbiosciences.com A Geno Technology, Inc. (USA) brand name PerfectFOCUS For Preparing Low Conductivity Samples for IEF/2D-Gel Electrophoresis
More informationCHAPTER 13: SOLUTIONS
CHAPTER 13: SOLUTIONS Problems: 1-8, 11-15, 20-30, 37-88, 107-110, 131-132 13.2 SOLUTIONS: HOMOGENEOUS MIXTURES solution: homogeneous mixture of substances present as atoms, ions, and/or molecules solute:
More informationISOLATE II PCR and Gel Kit. Product Manual
ISOLATE II PCR and Gel Kit Product Manual 2 Product Manual www.bioline.com/isolate PCR and Gel Kit ISOLATE II PCR and Gel Kit ISOLATE II PCR and Gel Kit 1 Kit contents 04 2 Description 04 3 Storage 04
More informationPRODUCTION OF MONOCLONAL ANTIBODIES FOR USE IN IMMUNOASSAYS BASED ON THE MAGNETIZABLE SOLID PHASE SEPARATION TECHNIQUE
PRODUCTION OF MONOCLONAL ANTIBODIES FOR USE IN IMMUNOASSAYS BASED ON THE MAGNETIZABLE SOLID PHASE SEPARATION TECHNIQUE W. CHAROENSIRIWATANA, N. JANEJAI, P.KRASAO XA9643133 Department of Medical Sciences,
More informationTIANquick Mini Purification Kit
TIANquick Mini Purification Kit For purification of PCR products, 100 bp to 20 kb www.tiangen.com TIANquick Mini Purification Kit (Spin column) Cat no. DP203 Kit Contents Contents Buffer BL Buffer PB Buffer
More informationDetermination of calcium by Standardized EDTA Solution
Determination of calcium by Standardized EDTA Solution Introduction The classic method of determining calcium and other suitable cations is titration with a standardized solution of ethylenediaminetetraacetic
More informationDairy Proteins. Table of Contents. Section Page. Cheese Milk Protein Review 2. Basic Flows and Definitions of Milk Products 4
Dairy Proteins This document, prepared by the Wisconsin Center for Dairy Research and the Wisconsin Milk Marketing Board, is intended to help clarify the present dairy protein issue. It can also be used
More informationPeptide Antibody Production
Peptide Antibody Production A) Peptide BioSynthesis (http://www.biosyn.com, 800-227-0627) B) Conjugation of peptide to KLH (Imject Maleimide Activated KLH, PIERCE=Thermo #77605, 10 mg) C) Peptide affinity
More informationChemistry B11 Chapter 6 Solutions and Colloids
Chemistry B11 Chapter 6 Solutions and Colloids Solutions: solutions have some properties: 1. The distribution of particles in a solution is uniform. Every part of the solution has exactly the same composition
More informationEnzyme Action: Testing Catalase Activity
Enzyme Action: Testing Catalase Activity Experiment 6A Many organisms can decompose hydrogen peroxide (H 2 O 2 ) enzymatically. Enzymes are globular proteins, responsible for most of the chemical activities
More informationGENOME RUSSIA PROJECT BLOOD SAMPLES COLLECTION, DNA EXTRACTION AND DNA QUALITY CONTROL PROTOCOLS
ST-PETERSBURG STATE UNIVERSITY THEODOSIUS DOBZHANSKY CENTER FOR GENOME BIOINFORMATICS GENOME RUSSIA PROJECT BLOOD SAMPLES COLLECTION, DNA EXTRACTION AND DNA QUALITY CONTROL PROTOCOLS 2015 The object of
More information50 g 650 L. *Average yields will vary depending upon a number of factors including type of phage, growth conditions used and developmental stage.
3430 Schmon Parkway Thorold, ON, Canada L2V 4Y6 Phone: 866-667-4362 (905) 227-8848 Fax: (905) 227-1061 Email: techsupport@norgenbiotek.com Phage DNA Isolation Kit Product # 46800, 46850 Product Insert
More informationEZ-Link NHS-Biotin Reagents
ITRUCTI EZ-Link -Biotin Reagents 3747. Meridian Road P.. Box 117 Rockford, IL 61105 20217 21336 21343 0237.4 umber Description 20217 EZ-Link -Biotin, 100 mg, -hydroxysuccinimidobiotin Molecular Weight:
More informationYour partner in immunology
Your partner in immunology Expertise Expertise Reactivity Reactivity Quality Quality Advice Advice Who are we? Specialist of antibody engineering Covalab is a French biotechnology company, specialised
More informationChromatin Immunoprecipitation (ChIP)
Chromatin Immunoprecipitation (ChIP) Day 1 A) DNA shearing 1. Samples Dissect tissue (One Mouse OBs) of interest and transfer to an eppendorf containing 0.5 ml of dissecting media (on ice) or PBS but without
More informationUltraClean PCR Clean-Up Kit
UltraClean PCR Clean-Up Kit Catalog No. Quantity 12500-50 50 Preps 12500-100 100 Preps 12500-250 250 Preps Instruction Manual Please recycle Version: 02212013 1 Table of Contents Introduction... 3 Protocol
More informationWorkshop 14-16 February 2006
Theoretical and practical approaches of Hepatocyte primary culture Workshop 14-16 February 2006 Lecture (2) Disaggregation & purification of target cells Coarse organizer Dr. Abo bakr Mohamed Eltayeb General
More informationChromatin Immunoprecipitation
Chromatin Immunoprecipitation A) Prepare a yeast culture (see the Galactose Induction Protocol for details). 1) Start a small culture (e.g. 2 ml) in YEPD or selective media from a single colony. 2) Spin
More informationTECHNICAL BULLETIN. FluoroTag FITC Conjugation Kit. Product Number FITC1 Storage Temperature 2 8 C
FluoroTag FITC Conjugation Kit Product Number FITC1 Storage Temperature 2 8 C TECHNICAL BULLETIN Product Description The FluoroTag FITC Conjugation Kit is suitable for the conjugation of polyclonal and
More informationInterim Progress Report R&D Project 348. Development of a Field Test Kit for Detection of Blue-Green Algal Toxins
Interim Progress Report R&D Project 348 Development of a Field Test Kit for Detection of Blue-Green Algal Toxins Biocode Limited November 1992 R&D 348/04/A ENVIRONMENT AGENCY 135357 CONTENTS SUMMARY KEYWORDS
More informationPART I: Neurons and the Nerve Impulse
PART I: Neurons and the Nerve Impulse Identify each of the labeled structures of the neuron below. A. B. C. D. E. F. G. Identify each of the labeled structures of the neuron below. A. dendrites B. nucleus
More informationTaking Apart the Pieces
Lab 4 Taking Apart the Pieces How does starting your morning out right relate to relief from a headache? I t is a lazy Saturday morning and you ve just awakened to your favorite cereal Morning Trails and
More informationAnalysis of Casein and Whey Protein in Whole, 2%, and Skim Milk by Capillary Gel Electrophoresis
Analysis of Casein and Whey Protein in Whole, 2%, and Skim Milk by Capillary Gel Electrophoresis Marcia Santos, Staff Applications Scientist, Beckman Coulter Life Sciences, Brea, CA USA Mark Lies, Marketing
More informationExperiment 3: Extraction: Separation of an Acidic, a Basic and a Neutral Substance
1 Experiment 3: Extraction: Separation of an Acidic, a Basic and a Neutral Substance Read pp 142-155, 161-162, Chapter 10 and pp 163-173, Chapter 11, in LTOC. View the videos: 4.2 Extraction (Macroscale);
More informationLaboratory 5: Properties of Enzymes
Laboratory 5: Properties of Enzymes Technical Objectives 1. Accurately measure and transfer solutions with pipettes 2. Use a Spectrophotometer to study enzyme action. 3. Properly graph a set of data. Knowledge
More informationTwo-Dimensional Gel Electrophoresis (2-DGE)
- Introduction - Sample preparation - First dimension: Isoelectric focusing - Second dimension: SDS-PAGE - Detection of protein spots: staining - Imaging analysis & 2D Gel databases - Spot handling: excision,
More informationRubisco; easy Purification and Immunochemical Determination
Rubisco; easy Purification and Immunochemical Determination Ulrich Groß Justus-Liebig-Universität Gießen, Institute of Plant Nutrition, Department of Tissue Culture, Südanlage 6, D-35390 Giessen e-mail:
More informationPHYSICAL SEPARATION TECHNIQUES. Introduction
PHYSICAL SEPARATION TECHNIQUES Lab #2 Introduction When two or more substances, that do not react chemically, are blended together, the result is a mixture in which each component retains its individual
More informationSize Exclusion Chromatography
Size Exclusion Chromatography TOYOPEARL Resins for SEC TOYOPEARL TOYOPEARL TOYOPEARL HW-55 TOYOPEARL HW-65 TOYOPEARL TOSOH BIOSCIENCE TOSOH BIOSCIENCE LLC 56 Keystone Drive Montgomeryville, PA 896-967
More informationBIOCHEMICAL FRACTIONATION OF CHLOROPLASTS
BIOCHEMICAL FRACTIONATION OF CHLOROPLASTS During the past thirty years, a series of protocols has been developed that permits the homogenization of tissues and the subsequent fractionation and purification
More informationTEPZZ 59 Z9ZA_T EP 2 592 090 A1 (19) (11) EP 2 592 090 A1 (12) EUROPEAN PATENT APPLICATION
(19) TEPZZ 9 Z9ZA_T (11) EP 2 92 090 A1 (12) EUROPEAN PATENT APPLICATION (43) Date of publication:.0.13 Bulletin 13/ (21) Application number: 11188776.6 (1) Int Cl.: C07K 9/00 (06.01) C07K 1/18 (06.01)
More informationLab 10: Bacterial Transformation, part 2, DNA plasmid preps, Determining DNA Concentration and Purity
Lab 10: Bacterial Transformation, part 2, DNA plasmid preps, Determining DNA Concentration and Purity Today you analyze the results of your bacterial transformation from last week and determine the efficiency
More informationDP419 RNAsimple Total RNA Kit. RNAprep pure Series. DP501 mircute mirna Isolation Kit. DP438 MagGene Viral DNA / RNA Kit. DP405 TRNzol Reagent
Overview of TIANGEN Products DP419 RNAsimple Total RNA Kit DP430 RNAprep pure Kit(For Cell/Bacteria) DP315/DP315-R TIANamp Virus DNA/RNA Kit DP431 RNAprep pure Kit (For Tissue) Silica-membrane Technology
More informationOne problem often faced in qualitative analysis is to test for one ion in a
Chemistry 112 Laboratory: Silver Group Analysis Page 11 ANALYSIS OF THE SILVER GROUP CATIONS Ag + Pb Analysis of a Mixture of Cations One problem often faced in qualitative analysis is to test for one
More informationRPCI 004 v.002 Staining Procedure For all Directly Conjugated Reagents (Whole Blood Method)
Immune Tolerance Network RPCI 004 v.002 Staining Procedure For all Directly Conjugated Reagents (Whole Blood Method) Author: Paul Wallace, Director, RPCI Laboratory of Flow Cytometry Approved by: Paul
More informationColligative Properties
CH302 LaBrake and Vanden Bout Colligative Properties PROBLEM #1: Give the molecular formula, the van t hoff factor for the following Ionic Compounds as well as guess the solubility of the compounds. If
More informationTRI Reagent Solution. A. Product Description. RNA / DNA / Protein Isolation Reagent Part Number AM9738 100 ml
TRI Reagent Solution RNA / DNA / Protein Isolation Reagent Part Number AM9738 100 ml A. Product Description TRI Reagent * solution is a complete and ready-to-use reagent for the isolation of total RNA
More informationCalculation of Molar Masses. Molar Mass. Solutions. Solutions
Molar Mass Molar mass = Mass in grams of one mole of any element, numerically equal to its atomic weight Molar mass of molecules can be determined from the chemical formula and molar masses of elements
More informationThe Huntington Library, Art Collections, and Botanical Gardens. How Sweet It Is: Enzyme Action in Seed Germination
The Huntington Library, Art Collections, and Botanical Gardens How Sweet It Is: Enzyme Action in Seed Germination Overview This experiment is intended to familiarize students with the macromolecule starch,
More informationThe basic division explosives for forensic purposes is shown in Fig. 1
Isolation, concentration and determination of post blast residua by liquid chromatography with DAD detector capillary electrophoresis with DAD detector Firearm fumes (gunshot residua, GSR) - various metallic
More informationACID-BASE TITRATIONS: DETERMINATION OF CARBONATE BY TITRATION WITH HYDROCHLORIC ACID BACKGROUND
#3. Acid - Base Titrations 27 EXPERIMENT 3. ACID-BASE TITRATIONS: DETERMINATION OF CARBONATE BY TITRATION WITH HYDROCHLORIC ACID BACKGROUND Carbonate Equilibria In this experiment a solution of hydrochloric
More informationserum protein and A/ G ratio
serum protein and A/ G ratio Blood plasma contains at least 125 individual proteins. Serum ( as contrasted with plasma) is deficient in those coagulation protein which are consumed during the process of
More informationCatalytic Activity of Enzymes
Catalytic Activity of Enzymes Introduction Enzymes are biological molecules that catalyze (speed up) chemical reactions. You could call enzymes the Builders and Do-ers in the cell; without them, life could
More informationINSTRUCTIONS 56-1190-98. Edition AC
Sephacryl S-100 High Resolution Sephacryl S-200 High Resolution Sephacryl S-300 High Resolution Sephacryl S-400 High Resolution Sephacryl S-500 High Resolution INSTRUCTIONS Sephacryl High Resolution chromatography
More informationEXPERIMENT # 3 ELECTROLYTES AND NON-ELECTROLYTES
EXPERIMENT # 3 ELECTROLYTES AND NON-ELECTROLYTES Purpose: 1. To investigate the phenomenon of solution conductance. 2. To distinguish between compounds that form conducting solutions and compounds that
More informationEfficient Multi-Well Protein Purification Strategies
Application Note PN 33576 Efficient Multi-Well Protein Purification Strategies Introduction Many tools and techniques are available today for protein purification. Development of a purification process
More informationIsolation of Caffeine from Tea
Isolation of Caffeine from Tea Introduction A number of interesting, biologically active compounds have been isolated from plants. Isolating some of these natural products, as they are called, can require
More informationCreatine Kinase Microplate Assay Kit User Manual
Creatine Kinase Microplate Assay Kit User Manual Catalog # CAK1045 Detection and Quantification of Creatine Kinase (CK) Activity in Urine, Serum, Plasma, Tissue extracts, Cell lysate, Cell culture media
More information