Marmara Üniversitesi Fen-Edebiyat Fakültesi Kimya Bölümü / Biyokimya Anabilim Dalı PURIFICATION AND CHARACTERIZATION OF PROTEINS

Save this PDF as:
 WORD  PNG  TXT  JPG

Size: px
Start display at page:

Download "Marmara Üniversitesi Fen-Edebiyat Fakültesi Kimya Bölümü / Biyokimya Anabilim Dalı PURIFICATION AND CHARACTERIZATION OF PROTEINS"

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

Announcements Columbus Day 10/8 No class, 10/9 BU Monday!

Announcements Columbus Day 10/8 No class, 10/9 BU Monday! Announcements Columbus Day 10/8 No class, 10/9 BU Monday! Make up section for Tuesday discussion: Wednesday 10/10, 5-6 pm, SAR 102 Monday Section: Tuesday 10/9, 10-11 am KCB 106 Wednesday Section: Wed.

More information

6 Characterization of Casein and Bovine Serum Albumin

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 information

HiPer Ion Exchange Chromatography Teaching Kit

HiPer 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 information

Protein Purification and Analysis

Protein Purification and Analysis Protein Purification and Analysis Numbers of genes: Humans ~40,000 genes Yeast ~6000 genes Bacteria ~3000 genes Solubility of proteins important for purification: 60-80% soluble, 20-40% membrane Some proteins

More information

TECHNICAL BULLETIN. HIS-Select Nickel Affinity Gel. Catalog Number P6611 Storage Temperature 2 8 C

TECHNICAL 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 information

DNA SPOOLING 1 ISOLATION OF DNA FROM ONION

DNA 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 information

CHROMATOGRAPHY Lab 3 INTRODUCTION

CHROMATOGRAPHY Lab 3 INTRODUCTION CHROMATOGRAPHY Lab 3 contributed by Mike Brundage CAUTIONS AND PITFALLS Your column is packed with Sephedex beads and is very fragile. Do not move or bump the column while running the experiment.. Be sure

More information

Why would you need to purify protein?

Why would you need to purify protein? Why would you need to purify protein? Methods for Working with Protein 1. Protein Isolation A. Selection of a protein source i. tissue and cell cultures (bacteria, yeast, mammalian, etc.) ii. genetically

More information

Genomic DNA Extraction Kit INSTRUCTION MANUAL

Genomic 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 information

ISOLATION OF CAFFEINE FROM TEA

ISOLATION 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 information

State what factors affect the rate of diffusion and explain how the rate is affected.

State what factors affect the rate of diffusion and explain how the rate is affected. Lab 4 Diffusion and Osmosis Lab Objectives Upon completion of this lab you should be able to do the following. Define and/or use properly the following terms: solute, solvent, concentration gradient, diffusion,

More information

Affinity Chromatography

Affinity Chromatography PR100-02 G-Biosciences 1-800-628-7730 1-314-991-6034 technical@gbiosciences.com A Geno Technology, Inc. (USA) brand name Affinity Chromatography Teacher s Guidebook (Cat. # BE-417) think proteins! think

More information

Guide to Reverse Phase SpinColumns Chromatography for Sample Prep

Guide 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 information

Plant Genomic DNA Extraction using CTAB

Plant 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 information

Classic Immunoprecipitation

Classic 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 information

TECHNICAL BULLETIN. TRI Reagent For processing tissues, cells cultured in monolayer or cell pellets. Catalog Number T9424 Store at room temperature.

TECHNICAL BULLETIN. TRI Reagent For processing tissues, cells cultured in monolayer or cell pellets. Catalog Number T9424 Store at room temperature. 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 information

Experiment #10: Liquids, Liquid Mixtures and Solutions

Experiment #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 information

Expt. 4: ANALYSIS FOR SODIUM CARBONATE

Expt. 4: ANALYSIS FOR SODIUM CARBONATE Expt. 4: ANALYSIS FOR SODIUM CARBONATE Introduction In this experiment, a solution of hydrochloric acid is prepared, standardized against pure sodium carbonate, and used to determine the percentage of

More information

Affi-Prep Protein A Matrix Instruction Manual

Affi-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 information

Benchtop Mitochondria Isolation Protocol

Benchtop 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 information

General western blot protocol. Guidance for running an efficient and accurate experiment

General western blot protocol. Guidance for running an efficient and accurate experiment blot protocol Guidance for running an efficient and accurate experiment Contents Introduction Solution and reagents Sample lysis Sample preparation Loading and running the gel Antibody staining Useful

More information

LAB 11 PLASMID DNA MINIPREP

LAB 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 information

Biology 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 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 information

Bio-Gel P Polyacrylamide Gel Instruction Manual

Bio-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 information

Purification of Plasmid DNA

Purification 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 information

Pure-IP Western Blot Detection Kit

Pure-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 information

Pharmaceutical Biotechnology. Recombinant DNA technology Western blotting and SDS-PAGE

Pharmaceutical 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 information

Protein extraction from Tissues and Cultured Cells using Bioruptor Standard & Plus

Protein 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 information

Corning Spin-X UF Concentrator Selection and Use Guide

Corning 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 information

Chapter 3 Contd. Western blotting & SDS PAGE

Chapter 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 information

Application Guide... 2

Application 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 information

Genomic DNA Purification Student Laboratory Manual

Genomic 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 information

Catalase. ***You will be working with hot water, acids and bases in this laboratory*** ****Use Extreme Caution!!!****

Catalase. ***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 information

Chapter 5, Lesson 3 Why Does Water Dissolve Salt?

Chapter 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 information

Purification by Recrystallization

Purification by Recrystallization Experiment 2 Purification by Recrystallization Objectives 1) To be able to select an appropriate recrystallizing solvent. 2) To separate and purify acetanilide by recrystallization. 3) To compare the melting

More information

Ann.wellhouse@TouchStoneScience.net 1. Enzyme Function

Ann.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 information

Cells Need to Exchange Materials with the Extracellular Fluid. Membrane Transport. Plasma Membrane. Cells Must Control Movements of Materials

Cells Need to Exchange Materials with the Extracellular Fluid. Membrane Transport. Plasma Membrane. Cells Must Control Movements of Materials Membrane Transport Chapter 6 Cells Need to Exchange Materials with the Extracellular Fluid Take in nutrients O 2 energy substrates building materials cofactors Dispose of wastes CO 2 Urea Cells Must Control

More information

Properties of Proteins

Properties of Proteins Properties of Proteins Experiment #8 Objective: To study chemical and physical properties of proteins from natural sources (egg and milk) and some chemical reactions of amino acid residues in these proteins,

More information

Exercise 5. Cell Fractionation: Isolation of Mammalian Red Blood Cell Plasma Membrane Using Differential Centrifugation

Exercise 5. Cell Fractionation: Isolation of Mammalian Red Blood Cell Plasma Membrane Using Differential Centrifugation R e p r i n t e d f r o m G a l l i k S., C e l l B i o l o g y O L M P a g e 1 Exercise 5. Cell Fractionation: Isolation of Mammalian Red Blood Cell Plasma Membrane Using Differential Centrifugation A.

More information

A) HCl C) 52 g KCl in 100 g water at 80ºC A) temperature of the solution increases B) supersaturated D) low temperature and high pressure D) KClO3

A) HCl C) 52 g KCl in 100 g water at 80ºC A) temperature of the solution increases B) supersaturated D) low temperature and high pressure D) KClO3 1. Which compound becomes less soluble in water as the temperature of the solution is increased? A) HCl B) 2. The solubility of O3(s) in water increases as the A) temperature of the solution increases

More information

RiboZol RNA Extraction Reagents

RiboZol 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 information

1. The lipid layer that forms the foundation of cell membranes is primarily composed of molecules called.

1. The lipid layer that forms the foundation of cell membranes is primarily composed of molecules called. Cell Membranes 1. The lipid layer that forms the foundation of cell membranes is primarily composed of molecules called. 2. Due to the repellent nature of the polar water molecules, the tails of the phospholipids

More information

PCR and Sequencing Reaction Clean-Up Kit (Magnetic Bead System) 50 preps Product #60200

PCR 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 information

Protein purification methods, a practical approach

Protein 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 information

Expression 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 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 information

ReadyPrep Protein Extraction Kit (Cytoplasmic/Nuclear)

ReadyPrep 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 information

EXTRACTION OF DNA FROM CALF THYMUS CELLS Revised 2/1/96 Introduction

EXTRACTION 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 information

Reading instructions to partitioning in aqueous two-phase systems

Reading 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 information

Preparation of frequently used solutions

Preparation 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 information

HighPure Maxi Plasmid Kit

HighPure 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 information

SODIUM CARBOXYMETHYL CELLULOSE

SODIUM 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 information

Factors Affecting Enzyme Activity

Factors 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 information

FIGURE 2.18. A. The phosphate end of the molecule is polar (charged) and hydrophilic (attracted to water).

FIGURE 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 information

Process of Science: Using Diffusion and Osmosis

Process 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 information

catalase 2H 2 O 2 (l) ----> 2H 2 O (l) + O 2 (g)

catalase 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 information

Dairy Proteins. Table of Contents. Section Page. Cheese Milk Protein Review 2. Basic Flows and Definitions of Milk Products 4

Dairy 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 information

OSMOSIS 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 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 information

Protocol v002 Page 1 of 7 Agencourt RNAdvance Cell v2 Total RNA Isolation from Cultured Cells

Protocol v002 Page 1 of 7 Agencourt RNAdvance Cell v2 Total RNA Isolation from Cultured Cells Page 1 of 7 Agencourt RNAdvance Cell v2 Total RNA Isolation from Cultured Cells Please refer to http://www.agencourt.com/technical for updated protocols and refer to MSDS instructions http://www.beckmancoulter.com/customersupport/msds/msds.asp

More information

STABILITY: TRI REAGENT is stable at 25 C for at least two years from the date of purchase (3).

STABILITY: 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 information

Keystone Study Guide Module A: Cells and Cell Processes

Keystone Study Guide Module A: Cells and Cell Processes Keystone Study Guide Module A: Cells and Cell Processes Topic 1: Biological Principles Cells and the Organization of Life Characteristics of Life all living things share the following characteristics:

More information

Methods of Grading S/N Style of grading Percentage Score 1 Attendance, class work and assignment 10 2 Test 20 3 Examination 70 Total 100

Methods 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 information

Western Blotting. Prepare samples:

Western 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 information

# 14 Synthesis of Salicylic Acid from Aspirin Tablets

# 14 Synthesis of Salicylic Acid from Aspirin Tablets # 14 Synthesis of Salicylic Acid from Aspirin Tablets Purpose: Acetyl salicylic acid is extracted from aspirin tablets, then is hydrolyzed to form another white solid, salicylic acid. Introduction: The

More information

Basics of SPE Technology & Mechanisms. Pieter Grobler, Sigma-Aldrich RSA

Basics 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

SOLUTIONS EXPERIMENT 13

SOLUTIONS EXPERIMENT 13 SOLUTIONS EXPERIMENT 13 OBJECTIVE The objective of this experiment is to demonstrate the concepts of concentrations of solutions and the properties of solution. Colloids will be demonstrated. EQUIPMENT

More information

TOTAL PROTEIN FIBRINOGEN

TOTAL 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 information

Separation of Mixtures by Paper Chromatography

Separation of Mixtures by Paper Chromatography Exercise 14 Page 1 Illinois Central College CHEMISTRY 130 Laboratory Section: Name: Separation of Mixtures by Paper Chromatography Objectives In this experiment we will effect a separation of a mixture

More information

--not necessarily a protein! (all proteins are polypeptides, but the converse is not true)

--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 information

UltraClean Soil DNA Isolation Kit

UltraClean 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 information

MagExtractor -Genome-

MagExtractor -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 information

LAB EXERCISE: Enzymes I Catechol Oxidase

LAB EXERCISE: Enzymes I Catechol Oxidase LAB EXERCISE: Enzymes I Catechol Oxidase Laboratory Objectives After completing this lab topic, you should be able to: 1. Define enzyme and describe the activity of enzymes in cells. 2. Differentiate competitive

More information

Bangs Laboratories, Inc.

Bangs Laboratories, Inc. Microsphere Specialists Bangs Laboratories, Inc. 9025 Technology Drive, Fishers, IN 46038-2886 Phone: 317-570-7020 Fax: 317-570-7034 E-mail: info@bangslabs.com Web site: www.bangslabs.com Tech. Note #

More information

Solutions & Colloids

Solutions & Colloids Chemistry 100 Bettelheim, Brown, Campbell & Farrell Ninth Edition Introduction to General, Organic and Biochemistry Chapter 6 Solutions & Colloids Solutions Components of a Solution Solvent: The substance

More information

Honors 227 Fall 2007 Laboratory with Ms. Clark. Enzymes, Reactions, Metabolism and Homeostasis

Honors 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 information

PerfectFOCUS. For Preparing Low Conductivity Samples for IEF/2D-Gel Electrophoresis. (Cat. # 786-124, 786-124T)

PerfectFOCUS. 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 information

CHAPTER 13: SOLUTIONS

CHAPTER 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 information

Molecular 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 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 information

Proteomics Workflows & Technologies

Proteomics Workflows & Technologies Proteomics Workflows & Technologies Proteomics "The analysis of the entire PROTEin complement expressed by a genome, or by a cell or tissue type." Wasinger VC et al, Electrophoresis 16 (1995) Proteomics

More information

ISOLATE II PCR and Gel Kit. Product Manual

ISOLATE 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 information

ANALYSIS OF WATER FOR CHLORIDE AND SULFATE IONS

ANALYSIS OF WATER FOR CHLORIDE AND SULFATE IONS Test Procedure for ANALYSIS OF WATER FOR CHLORIDE AND SULFATE IONS TxDOT Designation: Tex-619-J Effective Date: August 2005 1. SCOPE 1.1 Use this method to analyze water for chloride and sulfate ions to

More information

Colligative properties of biological liquids

Colligative properties of biological liquids Colligative properties of biological liquids Colligative properties are properties of solutions that depend on the number of molecules in a given volume of solvent and not on the properties (e.g. size

More information

RNAzol RT RNA Isolation Reagent

RNAzol RT RNA Isolation Reagent G e n e C o p o eia TM Expressway to Discovery RNAzol RT RNA Isolation Reagent Most effective reagent for isolation of total RNA and small RNA from samples of various origin Cat. No. E01010A User Manual

More information

TIANquick Mini Purification Kit

TIANquick 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 information

Chemistry B11 Chapter 6 Solutions and Colloids

Chemistry 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 information

PRODUCTION 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 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 information

Peptide Antibody Production

Peptide 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 information

Factors that Affect the Rate of Dissolving and Solubility

Factors that Affect the Rate of Dissolving and Solubility Dissolving Factors that Affect the Rate of Dissolving and Solubility One very important property of a solution is the rate of, or how quickly a solute dissolves in a solvent. When dissolving occurs, there

More information

Determination of calcium by Standardized EDTA Solution

Determination 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 information

50 g 650 L. *Average yields will vary depending upon a number of factors including type of phage, growth conditions used and developmental stage.

50 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 information

Enzyme Action: Testing Catalase Activity

Enzyme 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 information

GENOME RUSSIA PROJECT BLOOD SAMPLES COLLECTION, DNA EXTRACTION AND DNA QUALITY CONTROL PROTOCOLS

GENOME 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 information

EXPERIMENT # 3 ELECTROLYTES AND NON-ELECTROLYTES

EXPERIMENT # 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 information

Workshop 14-16 February 2006

Workshop 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 information

Name Matter Questions Date:

Name Matter Questions Date: Name Matter Questions Date: 1. Which substance has a definite shape and a definite volume at STP? 1) NaCl(aq) 2) Cl2(g) 3) CCl4( ) 4) AlCl3(s) 2. Which two particle diagrams represent mixtures of diatornic

More information

Your partner in immunology

Your 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 information

TRI Reagent Solution. A. Product Description. RNA / DNA / Protein Isolation Reagent Part Number AM9738 100 ml

TRI 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 information

Chromatin Immunoprecipitation (ChIP)

Chromatin 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 information

UltraClean PCR Clean-Up Kit

UltraClean 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 information

IDENTIFICATION OF AN UNKNOWN ORGANIC COMPOUND Classification Tests

IDENTIFICATION OF AN UNKNOWN ORGANIC COMPOUND Classification Tests IDENTIFICATION OF AN UNKNOWN ORGANIC COMPOUND Classification Tests In this experiment you will attempt to identify an organic unknown from a selected group of compounds from the class of alcohols, aldehydes,

More information

3.3 Cell Membrane (p. 81)

3.3 Cell Membrane (p. 81) Name Bio PreAP/GT 3.3 Cell Membrane (p. 81) Cell Membranes are composed of two phospholipid layers. The cell membrane, or the membrane, forms a boundary between a cell and the outside environment and controls

More information