Why would you need to purify protein?
|
|
|
- Valerie Payne
- 7 years ago
- Views:
Transcription
1 Why would you need to purify protein?
2 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 engineered - tagged proteins, over-expression B. Solubilization i. osmotic lysis - hypotonic solution swelling and bursting ii. French press - high pressure & small orifice iii. sonicator iv. homogenizer -tissue grinder, v. glass beads versus mortar & pestle vi. dounce -.
3
4 What properties of proteins can be used to separate and purify them from each other?
5 Methods for Working with Protein 2. Separation methods A. Properties that are used to separate proteins: i. charge ii. hydrophobicity iii. affinity iv. solubility & stability v. molecular weight B. differential centrifugation - S-100 versus S-30 C. precipitation/solubility i. salting in versus salting out solubility of a protein close to its pi versus the effect of salt interacting w/ solvent & not the protein ii. examples of a. ammonium sulfate precipitation b. PEI - poly(ethyleneimine)precipitation
6
7 S solubility of protein in salt solution S` - solubility of protein in pure water
8 Protein Solubility Salt-in At lower ionic strength, increased salt increase solubility Salt-Out At higher ionic strengths Increased salt concentrations causes the protein to precipitate out of solution Competition between the added salt with other dissolved solutes for solvation
9 Solute-solute interactions > solute-solvent interactions
10
11 Solubility of lactoglobulin depends on ph
12 Methods for Working with Protein 2. Separation methods D. chromatography - overall example i. ion exchange - cation vs. anion a. strong versus weak (effect of ph) b. counterion present is important c. types of gradients and their application linear, nonlinear, step
13
14
15
16
17
18
19 Methods for Working with Protein 2. Separation methods D. chromatography - overall example ii. affinity chromatography a. ligand based: glutathione covalent linked to resin GST fusion protein b. speciality dyes - Cibracon blue and others c. Immunoaffinity: epitope tags such as FLAG, V5, etc. d. DNA - general and specific DNAs e. others (example: heparin, hydroxyapatite)
20
21
22 Methods for Working with Protein 2. Separation methods D. chromatography iii. gel exclusion or gel filtration a. separation based on size b. exclusion volume c. different size limit materials iv. HIC or hydrophobic interaction chromatography compare to reverse phase chromatography v. types of resin cellulose, dextran, agarose, polyacrylamide, perfusion beads
23 Sequence Databases Some specifically for proteins and others for DNA
24 PubMed access to multiple databases BLAST search
25 Gel filtration or size exclusion chromatography What is the size of the protein of interest and the contaminating protein? How best to separate them?
26
27
28
29
30
31
32
33
34
35
36
37
38 Precautions or concerns might you have about maintaining your protein s activity What can you do to avoid these problems?
39 Methods for Working with Protein 3. Stabilization of protein A. Changing buffer i. dialysis ii. ultracentrifugation B. Concentrating protein helps to maintain active protein sometimes addition of a carrier protein may help addition of glycerol C. Inhibitors i. proteases ii. phosphatase inhibitors
40
41
42
43 Methods for Working with Protein 4. Protein Analysis A. Gel electrophoresis i. Discontinous gel a. polyacrylamide: ratio bisacrylamide/ acrylamide b. TEMED - free radical stablizer c. ammonium persulfate d. stacking gel ph 6.8, contains glycine pk2=9.78 e. running gel is ph 8.8, sample contains bromophenol blue
44 Methods for Working with Protein 4. Protein Analysis A. Gel electrophoresis ii. SDS-PAGE a. SDS forms a micelle around the polypeptide the size and charge of the micelle is approximately proportional to the size of the polypeptide b. denatures proteins c. protein stains: Coomassie blue, silver stains, fluorescent stains d. gradient gels
45
46
47
48 SDS-PAGE principle Glycine is an amino acid, and its charge property changes depend on the ph. In stacking gel (ph6.8), only a little amount of glycine is negatively charged, thus, glycine moves very slowly. SDS-bound proteins move much faster in low-density gel. Thus, proteins are stacked on the running front of glycine. In separation gel (ph8.8), glycine is charged and moves very fast. Proteins can move dependent on their size. Stacking gel 5%, ph6.8 Separation gel 6-20%, ph8.8-+
49
50
51 Methods for Working with Protein 4. Protein Analysis B. 2-Dimensional gels 1st dimension: isoelectric focusing tube gel format ampholytes to create an immobilized ph gradient 2nd dimension is usually SDS-PAGE
52 General Formula of Ampholytes Each ampholyte has a different pk and isoelectric point. Each ampholyte has a particular buffering capacity
53 Methods for Working with Protein 4. Protein Analysis C. Immunoblotting D. Gel-shift or EMSA (electrophoretic mobility gel shift assay) assays
54 Western blotting
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.
Approaches that can be used to study expression of specific proteins
Approaches that can be used to study expression of specific proteins Receptors and transporters Homogenate binding studies Receptor autoradiography Radiochemical Western blotting Immunohistochemistry/cytochemistry
EZ-Run Protein Gel Solution. EZ-Run Protein Standards. EZ-Run Gel Staining Solution. Traditional SDS-Page Reagents. Protein Electrophoresis
EZ-Run Protein Gel Solution EZ-Run Protein Standards EZ-Run Gel Staining Solution Traditional SDS-Page Reagents Protein Electrophoresis protein electrophoresis Introduction Sodium dodecyl sulfate polyacrylamide
Marmara Üniversitesi Fen-Edebiyat Fakültesi Kimya Bölümü / Biyokimya Anabilim Dalı PURIFICATION AND CHARACTERIZATION OF PROTEINS
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
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
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
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)
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
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
7 Electrophoresis. µ proportional to Q
7 Electrophoresis Objectives: A) To perform agarose gel electrophoresis of the proteins isolated in last week's experiment and B) to interpret the banding patterns produced by these proteins. Introduction:
Lecture 10 PROTEIN ISOLATION
Lecture 10 PROTEIN ISOLATION 1 Protein isolation Outlines: assay; homogenization; fractionation; centrifugation; quantitation; chromatography, electrophoresis; sequencing; people. 2 Nobel Prize Winners
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
ReadyPrep Protein Extraction Kit (Soluble/Insoluble) Instruction Manual. Catalog #163-2085
ReadyPrep Protein Extraction Kit (Soluble/Insoluble) Instruction Manual Catalog #163-2085 For technical service, call your local Bio-Rad office, or in the US, call 1-800-4BIORAD (1-800-424-6723) Table
Electrophoresis and Electroblotting of Proteins
Electrophoresis and Electroblotting of Proteins The purpose of the next lab exercises will be to study the relative amounts of β-actin in cells of the B-16 melanoma, liver and muscle of mice. Electrophoresis
--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
Chapter 14 SDS-PAGE. Objectives
SDS-PAGE This lab will introduce you to SDS-PAGE, a simple and inexpensive method for resolving proteins in complex mixtures. SDS-PAGE gels provide the starting materials for western blots and for some
2D gel Protocol. 2. Determining Protein Concentration of cell lysates
2D gel Protocol 1. Lysis and Protein Extraction from cells Prepare cell lysates with Trizol extraction by following Kathleen Lyons s protocol: AfCS Procedure Protocol PP00000155, Version 1, 05/12/03 (Ref.1).
Discontinuous native protein gel electrophoresis
Discontinuous native protein gel electrophoresis Michael Niepmann and Junfeng Zheng Institute of Biochemistry Friedrichstrasse 24 Faculty of Medicine, JustusLIebigUniversity 35392 Giessen, Germany In this
SDS-PAGE. (June 23, 2005)
SDS-PAGE (June 23, 2005) GATHER REAGENTS & MATERIALS 30% acrylamide/0.8% bisacrylamide (30:1) 4X Tris. Cl/SDS, ph 8.8 4X Tris. Cl/SDS, ph 6.8 Ammonium persulfate, 10% (Make fresh each time.) SDS electrophoresis
Biochemistry Lab SDS PAGE and Western blot General Instructions
Background When an electrical field is applied across a solution, the movement of the charged particles (proteins) is influenced not only by the charge but also the voltage, distance between electrodes,
Two-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,
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.
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
Aurum Ion Exchange Mini Kits and Columns. Instruction Manual
Aurum Ion Exchange Mini Kits and Columns Instruction Manual Catalog # 732-6710 Aurum AEX Mini Kits, 2 pk 732-6705 Aurum AEX Mini Kits, 10 pk 732-6706 Aurum AEX Mini Columns, 25 pk 732-6707 Aurum AEX Mini
ELECTROPHORESIS SLAB (THIN LAYER GEL) AND CAPILLARY METHODS. A. General Introduction
ELECTROPHORESIS SLAB (THIN LAYER GEL) AND CAPILLARY METHODS A. General Introduction Electrophoresis: a separation method based on differential rate of migration of charged species in an applied dc electric
Protein immunoblotting
Protein immunoblotting (Western blotting) Dr. Serageldeen A. A. Sultan Lecturer of virology Dept. of Microbiology SVU, Qena, Egypt seaas@lycos.com Western blotting -It is an analytical technique used to
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
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
Recombinant Enterokinase Kits
Table of Contents About the Kits...2 Description 2 Components 2 rek Cleavage...3 Small scale optimization 3 Scale-up 4 Monitoring cleavage 4 rek Capture...5 Capture buffer considerations 5 Monitoring rek
2D gel electrophoresis
2D gel electrophoresis Required solutions Cathode buffer: TRIS 7.6g Glycine 36g SDS 2.5g Fill up with ddwater to 250ml ESS (equilibration stock solution) SDS 2g Urea 36g EDTA 3mg 50 mm TRIS-HCl ph 6.8
Electrophoresis EXPERIMENT 4. Theory
( F EXPERIMENT 4 Electrophoresis Theory Basic Principles Electrophoresis is the process of migration of charged molecules through solutions in an applied electric field. Electrophoresis is often classified
Protein Purification. Nison Sattayasai Khon Kaen University Thailand. 1. Introduction. 2. Extraction of protein
1 Protein Purification Nison Sattayasai Khon Kaen University Thailand 1. Introduction There are many thousand kinds of proteins with different properties and functions in a cell. To study a protein, one
How to isolate proteins. Manju Kapoor
How to isolate proteins Manju Kapoor Background Numerous authoritative books, excellent reviews and articles have been written on this subject. While general methods for isolation and purification of proteins
Western Blotting: Mini-gels
Western Blotting: Mini-gels Materials a Protein Extraction Buffer (for callus or kernel), Solution Stock Final Volume Tris-HCl ph 80 1 M 200 mm 20 ml NaCl 4 M 100 mm 25 ml Sucrose 2 M 400 mm 20 ml EDTA
#01 Polyacrylamide Slab Gel Electrophoresis of Proteins Hassan Ahmad, University of Texas-Pan American, Edinburg, TX 78539
#01 Polyacrylamide Slab Gel Electrophoresis of Proteins Hassan Ahmad, University of Texas-Pan American, Edinburg, TX 78539 INTRODUCTION Description Electrophoresis involves the transport of charged molecules
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
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
Detection of proteins by lithium dodecyl sulphate polyacrylamide gel electrophoresis
Detection of proteins by lithium dodecyl sulphate polyacrylamide gel electrophoresis During electrophoretic measurements a mixture of compounds in solution is taken into a chamber, two electrodes are joined
Proteomics in Practice
Reiner Westermeier, Torn Naven Hans-Rudolf Höpker Proteomics in Practice A Guide to Successful Experimental Design 2008 Wiley-VCH Verlag- Weinheim 978-3-527-31941-1 Preface Foreword XI XIII Abbreviations,
Western Blotting. USA: proteintech@ptglab.com UK & Europe: europe@ptglab.com China: service@ptglab.com. www.ptglab.com
Western Blotting All steps are carried out at room temperature unless otherwise indicated. Recipes for all solutions highlighted bold are included at the end of the protocol. SDS-PAGE 1. Construct an SDS-PAGE
PROTOCOL 1850 Millrace Drive, Suite 3A Eugene, Oregon 97403 www.mitosciences.com
PROTOCOL Western Blotting Transfer and Detection Procedure 1850 Millrace Drive, Suite 3A Eugene, Oregon 97403 02-11 DESCRIPTION Western Blotting Transfer and Detection Procedure ADDITIONAL MATERIALS REQUIRED
Electrophoresis Guide
Novex Pre-Cast Gel Electrophoresis Guide Version B January 27, 2003 IM-1002 Novex Pre-Cast Gel Electrophoresis Guide General information and protocols for using Novex pre-cast gels www.invitrogen.com tech_service@invitrogen.com
Size Exclusion Chromatography
Size Exclusion Chromatography Size Exclusion Chromatography Instructors Stan Hitomi Coordinator Math & Science San Ramon Valley Unified School District Danville, CA Kirk Brown Lead Instructor, Edward Teller
Section XIII: Protein Separation in Agarose Gels
Section XIII: In This Section Introduction 196 Buffers for in Agarose 197 Casting Agarose Gels for 198 Preparation and Loading of Protein Samples 198 Optimal Voltage and Electrophoretic Times 199 Detection
Protein Expression and Analysis. Vijay Yajnik, MD, PhD GI Unit MGH
Protein Expression and Analysis Vijay Yajnik, MD, PhD GI Unit MGH Identify your needs Antigen production Biochemical studies Cell Biology Protein interaction studies including proteomics Structural studies
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,
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
Methods for Protein Analysis
Methods for Protein Analysis 1. Protein Separation Methods The following is a quick review of some common methods used for protein separation: SDS-PAGE (SDS-polyacrylamide gel electrophoresis) separates
METHOD USED TO EXTRACT TOTAL MUSCLE PROTEIN FOR WESTERN BLOT USING TRIS-EDTA BUFFER*
METHOD USED TO EXTRACT TOTAL MUSCLE PROTEIN FOR WESTERN BLOT USING TRIS-EDTA BUFFER* SOLUTIONS FOR SAMPLE EXTRACTION 1. Tris-EDTA Buffer, ph 8.3 1 liter 50 mm Tris 6.06 g 10 mm EDTA 3.72 g Adjust ph to
Recombinant Protein Expression & Purification -- Challenges & Solutions
Recombinant Protein Expression & Purification -- Challenges & Solutions Liyan Pang, Ph.D. liyan.pang@genscript.com Table of Contents 1 2 3 4 5 6 Choose Expression System Optimize Protein Expression Protein
In vitro analysis of pri-mirna processing. by Drosha-DGCR8 complex. (Narry Kim s lab)
In vitro analysis of pri-mirna processing by Drosha-DGCR8 complex (Narry Kim s lab) 1-1. Preparation of radiolabeled pri-mirna transcript The RNA substrate for a cropping reaction can be prepared by in
About the Kits...2 Description 2 Components 2. Factor Xa Cleavage...3 Small scale optimization 3 Scale-up 3 Monitoring cleavage 4
Table of Contents About the Kits...2 Description 2 Components 2 Factor Xa Cleavage...3 Small scale optimization 3 Scale-up 3 Monitoring cleavage 4 Factor Xa Capture...5 Capture buffer considerations 5
WESTERN BLOTTING - A BEGINNER S GUIDE
WESTERN BLOTTING - A BEGINNER S GUIDE Western blotting identifies with specific antibodies proteins that have been separated from one another according to their size by gel electrophoresis. The blot is
Ready Gel System RESOURCE GUIDE
Ready Gel System RESOURCE GUIDE Ready Gel System Resource Guide Exploring, discovering, and understanding the world around us - the biological systems interacting with the environment - is a process that
Protocol for Western Blotting
Protocol for Western Blotting Materials Materials used on Day 3 Protease inhibitor stock: 1 μg/μl pepstatin A in DMSO 200 μm leupeptin in OG Buffer 200 mm PMSF: Freshly made. Ex) 34.8 mg PMSF in 1 ml isopropanol
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
Charged interfaces. Formation of charged surfaces. Helmholz, Gouy- Chapman and Stern model of double layer.
Charged interfaces. Formation of charged surfaces. Helmholz, Gouy- Chapman and Stern model of double layer. Charged interfaces In polar solvents, most substances become charged: Preferential adsorption
Rapid GST Inclusion Body Solubilization and Renaturation Kit
Product Manual Rapid GST Inclusion Body Solubilization and Renaturation Kit Catalog Number AKR-110 FOR RESEARCH USE ONLY Not for use in diagnostic procedures Introduction Bacteria are widely used for His
About the Kits...2 Description 2 Components 2
Table of Contents About the Kits...2 Description 2 Components 2 Thrombin Cleavage...3 Small scale optimization 3 Scale-up 4 Factors that affect thrombin activity 4 Monitoring cleavage 5 Biotinylated Thrombin
AES Application Focus Isoelectric Focusing Page 1
AES Application Focus Isoelectric Focusing Page 1 Isoelectric focusing Adapted from Chapter 7, Gel Electrophoresis of Proteins, by David E. Garfin, Pages 197-268, in Essential Cell Biology, Volume 1: Cell
Introduction to Bioprocessing
Introduction to Bioprocessing Cambridge Healthtech Institute Peptalk Palm Springs, CA Presented by Susan Dana Jones and Sheila Magil BioProcess Technology Consultants www.bptc.com BioProcess Technology
Developing Protocols of Tricine-SDS-PAGE for Separation of Polypeptides in the Mass Range 1-30 kda with Minigel Electrophoresis System
Int. J. Electrochem. Sci., 11 (2016) 640-649 International Journal of ELECTROCHEMICAL SCIENCE www.electrochemsci.org Developing Protocols of Tricine-SDS-PAGE for Separation of Polypeptides in the Mass
Introduction to Proteomics
Introduction to Proteomics Åsa Wheelock, Ph.D. Division of Respiratory Medicine & Karolinska Biomics Center asa.wheelock@ki.se In: Systems Biology and the Omics Cascade, Karolinska Institutet, June 9-13,
Recombinant Protein Expression and Purification from E. coli
Recombinant Protein Expression and Purification from E. coli Many different biochemistry projects require recombinant protein. This is a simple method for creating crude bacterial lysate of IPTG inducible
A Practical Approach on SDS PAGE for Separation of Protein
A Practical Approach on SDS PAGE for Separation of Protein Suvra Roy, Vikash Kumar* Central Inland Fisheries Research Institute (CIFRI), Barrackpore, 700120, India Abstract: Polyacrylamide gel electrophoresis
Molecular Biology Techniques: A Classroom Laboratory Manual THIRD EDITION
Molecular Biology Techniques: A Classroom Laboratory Manual THIRD EDITION Susan Carson Heather B. Miller D.Scott Witherow ELSEVIER AMSTERDAM BOSTON HEIDELBERG LONDON NEW YORK OXFORD PARIS SAN DIEGO SAN
Immunoblotting (Western blotting)
Immunoblotting (Western blotting) HIV Transfer to NC membrane membrane gel support buffer Dissociate in SDS Incubate with 1ary antiserum Separate by SDS-PAG Incubate w/ labeled 2nd ab Immunoblotting :
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
Heterologous expression and purification of proteins in E. coli
Heterologous expression and purification of proteins in E. coli Rory Koenen Institut für molekulare Herz-Kreislaufforschung University Hospital of the RWTH Aachen rkoenen@ukaachen.de Tel. 35984 contents
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
SPE and HPLC. Dr Iva Chianella Lecturer in Analytical Chemistry Cranfield Health +44 (0) 1234 758322. i.chianella.1998@cranfield.ac.
SPE and HPLC Dr Iva Chianella Lecturer in Analytical Chemistry Cranfield Health +44 (0) 1234 758322 i.chianella.1998@cranfield.ac.uk Solid-Phase Extraction- SPE Simple, fast and efficient sample preparation
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
Microarray Technology
Microarrays And Functional Genomics CPSC265 Matt Hudson Microarray Technology Relatively young technology Usually used like a Northern blot can determine the amount of mrna for a particular gene Except
Definitive guide to western blot
Definitive guide to western blot Western blotting uses specific antibodies to identify proteins that have been separated based on size by gel electrophoresis. The immunoassay uses a membrane made of nitrocellulose
Glutathione Resin. User Manual. User Manual. Cat. Nos. 635607, 635608, 635619 PT3306-1 (071414)
User Manual Glutathione Resin User Manual United States/Canada 800.662.2566 Asia Pacific +1.650.919.7300 Europe +33.(0)1.3904.6880 Japan +81.(0)77.543.6116 Cat. Nos. 635607, 635608, 635619 PT3306-1 (071414)
Organic Molecules of Life - Exercise 2
Organic Molecules of Life - Exercise 2 Objectives -Know the difference between a reducing sugar and a non-reducing sugar. -Distinguish Monosaccharides from Disaccharides and Polysaccharides -Understand
Electrophoresis, cleaning up on spin-columns, labeling of PCR products and preparation extended products for sequencing
Electrophoresis, cleaning up on spin-columns, labeling of PCR products and preparation extended products for sequencing PAGE electrophoresis Polyacrylamide gel electrophoresis (PAGE) is used for separating
INSTRUCTIONS 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
Lab 4: Osmosis and Diffusion
Lab 4: Osmosis and Diffusion The plasma membrane enclosing every cell is the boundary that separates the cell from its external environment. It is not an impermeable barrier, but like all biological membranes,
DP419 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
EZ-PAGE Electrophoresis System USER MANUAL
EZ-PAGE Electrophoresis System USER MANUAL Table of Contents Safety Information.. 2 Product Description... 2 Product Contents..... 3 Specifications & Storage Conditions.. 3 Product Use..... 3 Getting Started
Expression. Labeling. Crystallization. Purification. 伯 森 生 物 科 技 股 份 限 公 司 Blossom Biotechnologies Inc. P. 8 P. 3 P. 5 P. 6 P. 6 P. 2 P. 7 P.
伯 森 蛋 白 質 研 究 產 品 專 刊 ProteinResearchProductCatalog Expression Purification Crystallization Labeling P. 2 LEXSY Protein Expression System P. 3 P. 5 P. 6 P. 6 Affinity Size Exclusion Ion Exchange FPLC Accessories
Protein transfer from SDS-PAGE to nitrocellulose membrane using the Trans-Blot SD cell (Western).
Western Blot SOP Protein transfer from SDS-PAGE to nitrocellulose membrane using the Trans-Blot SD cell (Western). Date: 8/16/05, 10/31/05, 2/6/06 Author: N.Oganesyan, R. Kim Edited by: R. Kim Summary:
RESOURCE Q, 1 ml and 6 ml RESOURCE S, 1 ml and 6 ml
GE Healthcare Life Sciences Instructions 71-7146-00 AI Ion Exchange Columns RESOURCE Q, 1 ml and 6 ml RESOURCE S, 1 ml and 6 ml Introduction RESOURCE Q and S are pre-packed columns for separating biomolecules
ELUTION OF DNA FROM AGAROSE GELS
ELUTION OF DNA FROM AGAROSE GELS OBTECTIVE: To isolate specific bands or regions of agarose-separated DNA for use in subsequent experiments and/or procedures. INTRODUCTION: It is sometimes necessary to
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
Chapter 3. Protein Structure and Function
Chapter 3 Protein Structure and Function Broad functional classes So Proteins have structure and function... Fine! -Why do we care to know more???? Understanding functional architechture gives us POWER
Transmembrane proteins span the bilayer. α-helix transmembrane domain. Multiple transmembrane helices in one polypeptide
Transmembrane proteins span the bilayer α-helix transmembrane domain Hydrophobic R groups of a.a. interact with fatty acid chains Multiple transmembrane helices in one polypeptide Polar a.a. Hydrophilic
RNA) - - - = 1 1 = 1 EU
Colorimetric Methods for Determining Protein Concentration. Goals: 1. Learn how to use colorimetric (Lowry, BCA, and Bradford) methods to determine protein concentration in mg/ml. 2. Use intrinsic biomolecular
Enzyme Activity and Assays
Robert K Scopes, La Trobe University, Bundoora, Victoria, Australia Enzyme activity refers to the general catalytic properties of an enzyme, and enzyme assays are standardized procedures for measuring
Standards for Electrophoresis and Blotting. A Wide Range for All Applications
for Electrophoresis and Blotting A Wide Range for All Applications A Wide Range of Bio-Rad s standards provide an excellent means of monitoring electrophoresis and blotting experiments. Bio-Rad has formulated
Chapter 6: Antigen-Antibody Interactions
Chapter 6: Antigen-Antibody Interactions I. Strength of Ag-Ab interactions A. Antibody Affinity - strength of total noncovalent interactions between single Ag-binding site on an Ab and a single epitope
using immobilized ph gradients
2-D Electrophoresis using immobilized ph gradients Principles and Methods 80-6429-60 Edition AC Handbooks from Amersham Biosciences Antibody Purification Handbook 18-1037-46 The Recombinant Protein Handbook
BUFFERS and MEDIAS Coomassie Blue Staining Solution Coomassie blue Destaining Solution DMEM Normal Cell Culture Media
BUFFERS and MEDIAS Coomassie Blue Staining Solution 2 g Coomassie Blue 2 L Methanol or Ethanol * 1.6 L 400 ml Glacial acetic acid *If you will be microwaving the gel in staining solution for rapid staining
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,
Nucleic Acid Techniques in Bacterial Systematics
Nucleic Acid Techniques in Bacterial Systematics Edited by Erko Stackebrandt Department of Microbiology University of Queensland St Lucia, Australia and Michael Goodfellow Department of Microbiology University
Chem 405 Biochemistry Lab I Experiment 2 Quantitation of an unknown protein solution.
Chem 405 Biochemistry Lab I Experiment 2 Quantitation of an unknown protein solution. Introduction: The determination of protein concentration is frequently required in biochemical work. Several methods
THE His Tag Antibody, mab, Mouse
THE His Tag Antibody, mab, Mouse Cat. No. A00186 Technical Manual No. TM0243 Update date 01052011 I Description.... 1 II Key Features. 2 III Storage 2 IV Applications.... 2 V Examples - ELISA..... 2 VI
BIOCHEMICAL 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
Recombinant Protein Purification Handbook
GE Healthcare Recombinant Protein Purification Handbook Principles and Methods ination at work imagination at work Handbooks from GE Healthcare Protein Purification Handbook 18-1132-29 Gel Filtration Principles