Fluorescent Difference Gel Electrophoresis
|
|
- Augusta Quinn
- 6 years ago
- Views:
Transcription
1 1 of 5 8/21/ :50 AM Please cite as: CSH Protocols; 2006; doi: /pdb.prot4233 Protocol Fluorescent Difference Gel Electrophoresis Angelika Görg, Oliver Drews, and Walter Weiss This protocol was adapted from "Fluorescent Difference Gel Electrophoresis," contributed by Angelika Görg, Oliver Drews, and Walter Weiss, Chapter 16, in Purifying Proteins for Proteomics (ed. Simpson). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, INTRODUCTION A bottleneck for high-throughput proteomic studies is image analysis. In conventional two-dimensional (2D) methodology, protein samples are separated on individual gels, stained, and quantified, followed by image comparison with computer-aided image analysis programs. Since different images are not always perfectly superimposable, image analysis is very time consuming. Fluorescent difference gel electrophoresis shortens this procedure (Ünlü et al. 1997) by independently labeling samples in vitro before IEF using two different fluorescent cyanine dyes, then mixing and separating the samples on a single 2D gel. Subsequent image analysis enables the differences (e.g., up- or down-regulated proteins) between the samples to be visualized. MATERIALS Reagents Cell or tissue sample for protein extraction CyDye DIGE Fluor, Cy2, Cy3, Cy5 minimal dye (Amersham Biosciences) Dimethylformamide (DMF) (99.8% anhydrous) IMPORTANT: The DMF should be fresh (<3 months old after first opening), because it degrades to amine compounds, which interfere with the labeling reaction. Dithiothreitol (DTT) solution [50% (w/v)] Prepare fresh before use! 2DQuant kit (Amersham Biosciences) ph indicator strips (ph ) Pharmalyte (ph 3-10) (Amersham Biosciences)
2 2 of 5 8/21/ :50 AM SDS sample solubilization buffer Lysine (10 mm) Thiourea/urea/CHAPS buffer Equipment Image analysis software (e.g., DeCyder Differential Analysis Software; Amersham Biosciences) Low-fluorescence glass plate (3 mm thick) Multicolor confocal laser scanner (e.g., Typhoon 9400) Software for image overlay and editing (e.g., ImageQuant Solution [included in Typhoon package]) METHOD 1. Extract proteins with SDS sample solubilization buffer (see Extraction and Solubilization of Total Protein from Microorganisms). Determine the protein concentration of the sample with the 2DQuant kit. Dilute the SDS extract with a fourfold excess of thiourea/urea/chaps buffer to a protein concentration of 1 µg/µl and 20 µg/µl (final SDS concentration must be 0.2%). Adjust the ph of the sample to 8.5 (±0.1). IMPORTANT: Primary amines such as carrier ampholytes, as well as reducing agents such as dithiothreitol (DTT), diminish the labeling efficiency. Therefore, the common urea lysis buffer cannot be used for protein extraction. For a comprehensive list of partially or completely incompatible compounds, see the CyDye manual (from Amersham Biosciences). Proteins with low cystine content can be solubilized directly in thiourea/urea/chaps buffer. 2. Reconstitute the CyDyes in DMF to a stock solution of 1 mm. See the CyDyes manual for a detailed protocol. 3. Directly before labeling, dilute the CyDye stock solution to a final concentration of 0.4 mm (400 pmol/µl); for example, mix 3 µl of DMF with 2 µl of reconstituted CyDye. 4. Prepare an aliquot of 50 µg of the protein sample, and add 400 pmol (1 µl) of the CyDye. Keep the dyes and the samples in the dark and on ice! 5. Vortex the sample briefly, and centrifuge the mixture in a microfuge at 10,000g for 5 seconds. 6. Incubate the sample in the dark for 30 minutes on ice. 7. Add 1 µl of 10 mm lysine to stop the labeling reaction. 8. Mix, centrifuge briefly, and incubate in the dark for 10 minutes on ice.
3 3 of 5 8/21/ :50 AM 9. Add 1 µl of DTT solution and 1 µl of Pharmalyte per 50 µl of sample solution. The final buffer concentration before isoelectric focusing (IEF) should not exceed 30 mm Tris. 10. Pool the protein samples that are going to be separated on the same first- and second-dimension gel, and immediately apply the mixture to the immobilized ph gradient (IPG) strip. Run IEF and SDS-PAGE as described in Isoelectric Focusing in Immobilized ph Gradient Strips Using the IPGphor Unit: Sample In-Gel Rehydration, Isoelectric Focusing in Immobilized ph Gradient Strips Using the IPGphor Unit: Sample Cup Loading, and Multiple SDS-PAGE on Vertical Electrophoresis Units. Approximately the same amount of fluorescence-labeled proteins should be loaded as would be needed for silver staining. Alternatively, store labeled sample for up to 3 months at 80ºC. Protect the IPGphor during the IEF and the buffer tank during SDS-PAGE from light to minimize photobleaching of the fluorescence dyes. 11. Following 2D gel electrophoresis, visualize the proteins using low-fluorescence glass plates. Directly scan the gels in the glass cassettes after the second dimension to ensure that all gels have the same dimensions; doing so simplifies spot matching of different gels. The exterior of the glass plates must be carefully cleaned with H 2 O and dried with a lint-free laboratory wipe before the gel cassette is positioned on the scanner. Each fluorescent dye should be consecutively excited to avoid fluorescence cross-talk and scanned at a resolution of at least 200 µm with a proper filter. After the scan, software such as ImageQuant Solution easily provides an initial image overlay of the scanned channels of one gel, thus giving a quick overview of differences between the labeled extracts. It should be kept in mind that the protein patterns of one gel might differ slightly in protein concentration and background. Although the DIGE system is highly reliable in terms of quantitation, especially when an internal standard is included in the experimental design, biological variances or variances during the protein extraction might affect samples. Therefore, at least three gels of three individual protein extracts per experimental condition (e.g., control and stress) should be run, normalized, and compared before qualitative and quantitative changes can be considered significant. REFERENCES Ünlü M., Morgan M.E., Minden J.S Difference gel electrophoresis: A single gel method for detecting changes in protein extracts. Electrophoresis 18: [Medline] Caution Dimethylformamide (DMF) Dimethylformamide (DMF; also known as N,N-dimethylformamide) HCON(CH 3 ) 2 is irritating to the eyes, skin, and mucous membranes. It can exert its toxic effects through inhalation, ingestion, or skin absorption. Chronic inhalation can cause liver and kidney damage. Wear appropriate gloves and safety glasses. Use in a chemical fume hood.
4 4 of 5 8/21/ :50 AM Caution Dithiothreitol (DTT) Dithiothreitol (DTT) is a strong reducing agent that emits a foul odor. It may be harmful by inhalation, ingestion, or skin absorption. When working with the solid form or highly concentrated stocks, wear appropriate gloves and safety glasses and use in a chemical fume hood. Recipe SDS sample solubilization buffer 1% (w/v) SDS 100 mm Tris-HCl (ph 9.5) To prepare 50 ml of SDS sample buffer, dissolve 0.5 g of SDS and 0.6 g of Tris base in ~40 ml of deionized H 2 O. Titrate to ph 9.5 with 4 N HCl, filter, and adjust the volume to 50 ml with deionized H 2 O. Recipe Thiourea/urea/CHAPs buffer 2 M thiourea 7 M urea 4% (w/v) CHAPS To prepare 50 ml of buffer, dissolve 22.0 g of urea and 8.0 g of thiourea in ~25 ml of deionized H 2 O. Add 0.5 g of Serdolit MB-1 ion-exchange resin (Serva), stir for 10 minutes, and filter. Add 2.0 g of CHAPS to the filtrate and adjust the final volume to 50 ml with deionized H 2 O. Copyright 2006 by Cold Spring Harbor Laboratory Press. Online ISSN: Terms of Service All rights reserved. Anyone using the procedures outlined in these protocols does so at their own risk. Cold Spring Harbor Laboratory makes no representations or warranties with respect to the material set forth in these protocols and has no liability in connection with their use. All materials used in these protocols, but not limited to those highlighted with the Warning icon, may be considered hazardous and should be used with caution. For a full listing of cautions, click here. All rights reserved. No part of these pages, either text or images, may be used for any reason other than personal use. Reproduction, modification, storage in a retrieval system or retransmission, in any form or by any means-electronic, mechanical, or otherwise-for reasons other than personal use is strictly prohibited without prior written permission.
5 5 of 5 8/21/ :50 AM CiteULike Connotea Del.icio.us Digg Reddit Technorati What's this?
Protocol INTRODUCTION MATERIALS. Directional Cloning of PCR Products
1 of 9 4/29/2009 3:49 PM Cite as: Cold Spring Harb. Protoc.; 2006; doi:10.1101/pdb.prot4140 Protocol Directional Cloning of PCR Products Gina L. Costa and Michael P. Weiner This protocol was adapted from
More informationProtocol INTRODUCTION RELATED INFORMATION MATERIALS. Histological Preparation of Embryonic and Adult Zebrafish Eyes
Please cite as: CSH Protocols; 2007; doi:10.1101/pdb.prot4846 Protocol Histological Preparation of Embryonic and Adult Zebrafish Eyes Richard J. Nuckels 1 and Jeffrey M. Gross 1,2,3,4 1 Section of Molecular
More informationLecture 13: Analysis of 2D gels
Lecture 13: Analysis of 2D gels A complete proteomic analysis aims at collecting quantitative information about all protein in a sample. A normal 2 Dimensional gel electrophoresis results are analyzed
More information4. The Second-dimensional SDS-PAGE (vertical) Protocol
4. The Second-dimensional SDS-PAGE (vertical) Protocol I. PURPOSE This procedure outlines the steps that must be carried out in the seconddimension SDS-PAGE using vertical system. II. ENVIRONMENT All work
More informationelectrophoresis tech Versatile Separation Capabilities of the PROTEAN i12 IEF System
electrophoresis tech note 6139 Versatile Separation Capabilities of the PROTEAN i12 IEF System ChengJun Sun, Adriana Harbers, Tom Berkelman, and Katy McGirr, Life Science Group, Bio-Rad Laboratories Inc.,
More informationAbsolutely RNA Microprep Kit
Absolutely RNA Microprep Kit INSTRUCTION MANUAL Catalog #400805 Revision A For In Vitro Use Only 400805-12 LIMITED PRODUCT WARRANTY This warranty limits our liability to replacement of this product. No
More informationPreAnalytiX Supplementary Protocol
PreAnalytiX Supplementary Protocol Purification of total RNA from microdissected PAXgene Tissue fixed, paraffin-embedded (PFPE) and PAXgene Tissue fixed, cryo-embedded (PFCE) tissues This protocol is for
More informationGE Healthcare. 2-D Electrophoresis. Principles and Methods
GE Healthcare 2-D Electrophoresis Principles and Methods Handbooks from GE Healthcare Antibody Purification Handbook 18-1037-46 The Recombinant Protein Handbook Protein Amplification and Simple Purification
More informationby nexttec TM 1 -Step
Protocol DNA Isolation from Tissue & Cells by nexttec TM 1 -Step - nexttec cleancolumns - Cat. No. 10N.010 Cat. No. 10N.050 Cat. No. 10N.250 Version 1.0 For research only Principle nexttec 1 -Step is the
More informationGE Healthcare. 2-D Electrophoresis. Principles and Methods
GE Healthcare 2-D Electrophoresis Principles and Methods Introduction Introduction to this handbook This handbook is intended to provide guidelines for performing high-resolution two-dimensional (2-D)
More informationLIFluor EnhanCE. for use with dsdna 1000 and dsdna 20,000 Kits
LIFluor EnhanCE for use with dsdna 1000 and dsdna 20,000 Kits For the Separation of Double Stranded DNA by P/ACE Fluorescent Capillary Electrophoresis 725824 AB September 2006 Beckman Coulter, Inc. 4300
More informationTHE INSTITUTE FOR GENOMIC RESEARCH Standard Operating Procedure SOP #: M004 REVISION LEVEL:.3 EFFECTIVE DATE: 9/16/03
Standard Operating Procedure PAGE: 1 of 8 SOP #: M004 REVISION LEVEL:.3 EFFECTIVE DATE: 9/16/03 AUTHOR: Jeremy Hasseman PRIMARY REVIEWERS: Renee Gaspard, Bryan Frank 1. PURPOSE This protocol describes
More informationDepartment of Microbiology, Lab 016 instructions
Protocol for standard FISH and DOPE-FISH for prokaryotes (slightly modified from Amann, 1995, note, for other modifications or other microorganisms like eukaryotes, consult special literature or check
More informationStrataPrep Plasmid Miniprep Kit
StrataPrep Plasmid Miniprep Kit INSTRUCTION MANUAL Catalog #400761 and #400763 Revision A For In Vitro Use Only 400761-12 LIMITED PRODUCT WARRANTY This warranty limits our liability to replacement of this
More informationGeneaid DNA Isolation Kit (Yeast)
Geneaid DNA Isolation Kit (Yeast) GEY100, GEY300 Advantages Sample: up to 2 10 8 yeast and other fungus species Yield: high yield, high quality DNA (A260/A280 = 1.8-2.0) Format: scalable DNA precipitation
More informationby nexttec TM 1 -Step
Special protocol DNA Isolation Drosophila by nexttec TM 1 -Step - nexttec cleanplate96 - Cat. No. 10N.901 Cat. No. 10N.902 Cat. No. 10N.904 Cat. No. 10N.924 Version 1.0 For research only Principle nexttec
More informationMICROARRAY HYBRIDISATION PROTOCOL 10/05 (for both PCR and Oligo arrays on UltraGAPS slides)
MICROARRAY HYBRIDISATION PROTOCOL 10/05 (for both PCR and Oligo arrays on UltraGAPS slides) Vassilis Mersinias, Giselda Bucca and Graham Hotchkiss Quick Protocol (see notes at end for detailed instructions)
More informationNoviPure Microbial Protein Kit (50)
Saving You Time For Life NoviPure Microbial Protein Kit (50) Catalog No. 47044 Quantity: 50 preps INSTRUCTION MANUAL Version 04202016 Please recycle www.mobio.com T: 800-606-6246 T: 760-929-9911 technical@mobio.com
More informationComplete kit for the systematic analysis of protein kinases from a proteome such as cell lysate, tissue or microorganisms.
Protein Kinase Landscape (Large Format PEP) Catalog # AB000504 Complete kit for the systematic analysis of protein kinases from a proteome such as cell lysate, tissue or microorganisms. Please read this
More informationEarlyTox Cell Integrity Kit
EarlyTox Cell Integrity Kit The EarlyTox Cell Integrity Kit from Molecular Devices is an optimized set of reagents that simplifies the measurement of live and dead cells in a single well. The assay uses
More informationTruePrime Single Cell WGA Kit
TruePrime SINGLE CELL WGA KIT HANDBOOK TruePrime Single Cell WGA Kit INDEX Legal...4 Intended use...4 Kit contents...5 Shipping and storage...5 Handling...6 Quality control...6 Reagents and equipment to
More informationPreAnalytiX Supplementary Protocol
PreAnalytiX Supplementary Protocol Purification of full-length proteins from sections of PAXgene Tissue fixed, paraffin-embedded (PFPE) tissue This protocol describes using the Qproteome FFPE Tissue kit
More informationProtocol for DNA extraction from FFPE Samples
25, ave Georges Lemaître - B-6041 Gosselies (Belgique) Tel : +32 (0)71 37 85 27 - Fax : +32 (0)71 34 78 79 Protocol for DNA extraction from FFPE Samples Step 1. Paraffin Removal 1 Equilibrate a heat block
More informationAnaTag HiLyte Fluor 647 Protein Labeling Kit
AnaTag HiLyte Fluor 647 Protein Labeling Kit Catalog # 72049 Kit Size 3 Conjugation Reactions This kit is optimized to conjugate HiLyte Fluor 647 SE to proteins (e.g., IgG). It provides ample materials
More informationNote: for laboratory research use only. RNA High-purity Total RNA Rapid Extraction Kit (Spin-column) Signalway Biotechnology
Note: for laboratory research use only RNA High-purity Total RNA Rapid Extraction Kit (Spin-column) Cat. #: RP1202 (50preps) Signalway Biotechnology I. Kit Content, Storage Condition and Stability Content
More informationssdna 100-R Kit Care and Use Instructions AG June 2015
Care and Use Instructions 726479 AG June 2015 Beckman Coulter, Inc. 250 S. Kraemer Blvd., Brea, CA 92821 USA 2015 Beckman Coulter, Inc. Copyright, Licenses and Trademarks Copyright Beckman Coulter, Inc.,
More informationSERVAGel IEF 3-10 Precast Vertical Gels for Isoelectric Focusing
INSTRUCTION MANUAL SERVAGel IEF 3-10 Precast Vertical Gels for Isoelectric Focusing (Cat. No. 43240, 43242) SERVA Electrophoresis GmbH Carl-Benz-Str. 7 D-69115 Heidelberg Phone +49-6221-138400, Fax +49-6221-1384010
More informationE.Z.N.A. MicroElute Genomic DNA Kit. D preps D preps D preps
E.Z.N.A. MicroElute Genomic DNA Kit D3096-00 5 preps D3096-01 50 preps D3096-02 200 preps December 2013 E.Z.N.A. MicroElute Genomic DNA Kit Table of Contents Introduction...2 Kit Contents/Storage and Stability...3
More informationPowerSoil DNA Isolation Kit
PowerSoil DNA Isolation Kit Catalog No. Quantity 12888-50 50 Preps 12888-100 100 Preps Instruction Manual Introduction The PowerSoil DNA Isolation Kit* is comprised of a novel and proprietary method for
More informationEZ-10 SPIN COLUMN GENOMIC DNA MINIPREPS KIT HANDBOOK
EZ-0 SPIN COLUMN GENOMIC DNA MINIPREPS KIT HANDBOOK (Bacteria, Plant, Animal, Blood) Version 8 Rev 05/0/03 EZ-0 Genomic DNA Kit Handbook Table of Contents Introduction Limitations of Use Features Applications
More informationInstant-Bands Protein Sample Loading Buffer for SDS-PAGE. User s Manual. View Protein Bands in an SDS Gel. Instantly. EZBiolab.
Instant-Bands Protein Sample Loading Buffer for SDS-PAGE User s Manual View Protein Bands in an SDS Gel Instantly www.ezbiolab.com 2 Instant-Bands User s Manual Table of Contents Introduction 3 Storage
More informationab Cy3 Fast Conjugation Kit
ab188287 Cy3 Fast Conjugation Kit Instructions for Use For the Covalent Conjugation of Antibodies or Proteins to Cy3 This product is for research use only and is not intended for diagnostic use. Version
More informationStellaris RNA FISH Protocol for Simultaneous IF + FISH in Adherent Cells
Stellaris RNA FISH Protocol for Simultaneous IF + FISH in Adherent Cells General Protocol & Storage Product Description A set of Stellaris RNA FISH Probes is comprised of up to 48 singly labeled oligonucleotides
More informationPinpoint Slide DNA Isolation System Catalog No. D3001
INSTRUCTIONS Pinpoint Slide DNA Isolation System Catalog No. D3001 Highlights Easily isolates genomic DNA in any targeted microscopic tissue area on a slide. The simple procedure combines Pinpoint tissue
More informationPROTOCOL 1850 Millrace Drive, Suite 3A Eugene, Oregon
PROTOCOL Pyruvate Dehydrogenase (PDH) Enzyme Activity Dipstick Assay Kit 1850 Millrace Drive, Suite 3A Eugene, Oregon 97403 MSP30 Rev.4 DESCRIPTION Pyruvate Dehydrogenase (PDH) Enzyme Activity Dipstick
More informationSample Preparation Demonstrated Protocol
10x Genomics Sample Preparation Demonstrated Protocol DNA Extraction from Fresh Frozen Tissue NOTICES Notices Manual Part Number CG000072 Rev B Legal Notices 2017 10x Genomics, Inc. All rights reserved.
More informationab Histone Extraction Kit
ab113476 Histone Extraction Kit Instructions for Use For the extraction of histone proteins from mammalian cells and tissue This product is for research use only and is not intended for diagnostic use.
More informationAmpliScribe T 7 Aminoallyl-RNA Transcription Kit
Cat. No. AA50125 The AmpliScribe T7 Aminoallyl-RNA Transcription Kit enables high-yield production of aminoallyl-labeled RNA. The kit utilizes Epicentre s high yielding AmpliScribe T7-Flash in vitro transcription
More informationProduct specification
Product specification FluoroLink TM Cy3-UTP PA 53026 Cy3 aminoallyl-utp Pre-availability use only Please note that: 1. Whilst the product has been tested before despatch, it has not been fully validated
More informationCode Description Molecular Weight Separation Range
SPRINT NEXT GEL * A Ready-to-Pour Acrylamide Gel for the Rapid Electrophoresis of Proteins Code Description Molecular Weight Separation Range Size M312-100ML M312-500ML M311-100ML M311-500ML SPRINT NEXT
More informationPresto Mini gdna Bacteria Kit
Instruction Manual Ver. 02.10.17 For Research Use Only Presto Mini gdna Bacteria Kit Advantages GBB004 (4 Preparation Sample Kit) GBB100/101 (100 Preparation Kit) GBB300/301 (300 Preparation Kit) Sample:
More informationab Optiblot Fluorescent Western Blot Kit
ab133410 Optiblot Fluorescent Western Blot Kit Instructions for Use For quantitative, multi-color fluorescent Western blotting. This product is for research use only and is not intended for diagnostic
More informationKit for total DNA isolation from yeast
Cat. No. EM10 Version: 1.2013 Kit for total DNA isolation from yeast EXTRACTME is a registered trademark of BLIRT S.A. www.dnagdansk.com Cat. No. EM10 I. INTENDED USE The EXTRACTME DNA YEAST kit is designed
More informationISOLATE II Blood DNA Kit. Product Manual
ISOLATE II Blood DNA Kit Product Manual 2 Product Manual www.bioline.com/isolate Blood DNA Kit ISOLATE II Blood DNA Kit ISOLATE II Blood DNA Kit 1 Kit contents 04 2 Description 04 3 Storage 05 4 Safety
More informationThe preparation of native chromatin from cultured human cells.
Native chromatin immunoprecipitation protocol The preparation of native chromatin from cultured human cells. All solutions need to be ice cold. Sucrose containing solutions must be made up fresh on the
More informationProtocol. Path-ID Multiplex One-Step RT-PCR Kit Protocol TaqMan probe-based multiplex one-step real-time RT-PCR detection of RNA targets
Protocol TaqMan probe-based multiplex one-step real-time RT-PCR detection of RNA targets For Research Use Only. Not for use in diagnostic procedures. Information in this document is subject to change without
More informationINOS. Colorimetric Cell-Based ELISA Kit. Catalog #: OKAG00807
INOS Colorimetric Cell-Based ELISA Kit Catalog #: OKAG00807 Please read the provided manual entirely prior to use as suggested experimental protocols may have changed. Research Purposes Only. Not Intended
More informationThermo Scientific GeneJET NGS Cleanup Kit
PRODUCT INFORMATION Thermo Scientific GeneJET NGS Cleanup Kit #K0851, #K0852 www.thermoscientific.com/onebio #K0851, K0852 Lot Expiry Date _ CERTIFICATE OF ANALYSIS Thermo Scientific GeneJET NGS Cleanup
More informationfoodproof Sample Preparation Kit III
For food testing purposes FOR IN VITRO USE ONLY Version 1, June 2015 For isolation of DNA from raw material and food products of plant and animal origin for PCR analysis Order No. S 400 06.1 Kit for 50
More informationWorking Reagent Preparation 1:1,000 dilution in standard Alcoholic Ortho- Phosphoric acid staining solution
www.smobio.com Product Information FluoroStain Protein Fluorescent Staining Dye (Red, 1,000X) PS1000 1 ml x 1 PS1001 1 ml x 5 Storage Protected from light -20 C for 24 months Working Reagent Preparation
More informationAbsolutely RNA Nanoprep Kit
Absolutely RNA Nanoprep Kit INSTRUCTION MANUAL Catalog #400753 Revision A For In Vitro Use Only 400753-12 LIMITED PRODUCT WARRANTY This warranty limits our liability to replacement of this product. No
More informationCalcein AM Cell Viability Kit
Instructions For Research Use Only. Not For Use In Diagnostic Procedures Calcein AM Cell Viability Kit Catalog# 4892-010-K 1000 Tests* * Calculated based on using 1 μm final concentration of Calcein AM;
More informationE.Z.N.A. Blood DNA Mini Kit. D preps D preps D preps
E.Z.N.A. Blood DNA Mini Kit D3392-00 5 preps D3392-01 50 preps D3392-02 200 preps January 2017 E.Z.N.A. Blood DNA Mini Kit Table of Contents Introduction and Overview...2 Illustrated Protocol...3 Kit Contents/Storage
More informationTECHNICAL BULLETIN. in vitro Translation Labeling System. FluoroTect Green Lys. Instruc ons for Use of Product L5001. Revised 11/13 TB285
TECHNICAL BULLETIN in vitro Translation Labeling System Instruc ons for Use of Product L5001 Revised 11/13 TB285 in vitro Translation Labeling System All technical literature is available at: www.promega.com/protocols/
More informationAmersham Western blotting
Amersham Western blotting Selection guide www.gelifesciences.com Innovative solutions designed to meet your specific protein analysis needs Since 1990 when we first launched Amersham ECL we have continued
More informationZR-96 DNA Sequencing Clean-up Kit Catalog Nos. D4052 & D4053
INSTRUCTION MANUAL ZR-96 DNA Sequencing Clean-up Kit Catalog Nos. D4052 & D4053 Highlights Simple 10 Minute Bind, Wash, Elute Procedure Flexible 15-20 µl Elution Volumes Allow for Direct Loading of Samples
More informationab R-Phycoerythrin Conjugation Kit Protocol
ab102918 R-Phycoerythrin Conjugation Kit Protocol Instructions for Use Antibody and protein modification This product is for research use only and is not intended for diagnostic use. Version 7 Last Updated
More informationcelldatasci.com/rnastorm RNAstorm RNA Isolation Kit for FFPE Tissue Samples Sample Kit (20 extractions)
celldatasci.com/rnastorm info@celldatasci.com RNAstorm RNA Isolation Kit for FFPE Tissue Samples Sample Kit (20 extractions) Support: Email: support@celldatasci.com Phone: 650.285.2376 (option 2) Toll-free:
More informationMATERIAL DATA SHEET. NOTE: Kit contains reagents sufficient for 10 x 30 μl reactions and 5 Western Blots (minigel. Reagents Provided in Kit
Lot # XXXXX ITCH/AIP4 Ubiquitin Ligase Kit Cat. # K-270 MATERIAL DATA SHEET The mammalian Itchy homolog, or ITCH, (also known as Atrophin-1-interacting protein 4 or AIP4) is a HECT domain class ubiquitin
More informationGenUP Virus DNA/RNA Kit
Product Insert GenUP Virus DNA/RNA Kit LOT: See product label EXPIRY DATE: See product label ORDERING INFORMATION PRODUCT GenUP Virus DNA/RNA Kit CAT. NO. BR0701101 BR0701102 BR0701103 SIZE 10 preps 50
More informationPureLink RNA Micro Kit
PureLink RNA Micro Kit For purification of total RNA from variety of samples in low elution volume Catalog no. 12183-016 Version B 19 December 2008 100003660 User Manual Table of Contents Kit Contents
More informationSDS PAGE. Reagents and Equipment. SERVA SDS PAGE Gels. Electrophoresis Buffers, Ready-to-Use. Reagents. Gel Media. SERVA Protein Standards
SDS PAGE Reagents and Equipment SERVA SDS PAGE Gels Electrophoresis Buffers, Ready-to-Use Reagents Gel Media SERVA Protein Standards SERVA Stains for SDS PAGE SDS PAGE Equipment All you need for... SDS
More informationLambda DNA Purification Kit
Lambda DNA Purification Kit INSTRUCTION MANUAL Catalog #200391 and #200392 Revision A For In Vitro Use Only 200391-12 LIMITED PRODUCT WARRANTY This warranty limits our liability to replacement of this
More informationOrexin A (HUMAN, MOUSE, RAT, PORCINE, OVINE,
Orexin A (HUMAN, MOUSE, RAT, PORCINE, OVINE, BOVINE) Western Blot Kit Protocol (Catalog #WBK-003-30) PHOENIX PHARMACEUTICALS, INC. TABLE OF CONTENTS 1. Kit Contents...2 2. Storage...2 3. Introduction...3
More informationVisualization of proteins electro-blotted on Hybond ECL and Hybond-P using Deep Purple Total Protein Stain
application note 2-D Electrophoresis Visualization of proteins electro-blotted on Hybond ECL d Hybond-P using Deep Purple Total Protein Stain S key words: Hybond ECL nitrocellulose Hybond-P polyvinylidene
More informationHiPer Gel Extraction Teaching Kit (Column Based)
HiPer Gel Extraction Teaching Kit (Column Based) Product Code: HTBM010 Number of experiments that can be performed: 10 Duration of Experiment Agarose Gel Electrophoresis: 1 hour Protocol: 1 hour Agarose
More informationInstructions for use
Instructions for use SARCOPTES-ELISA 2001 DOG Test to detect the presence of antibodies against the cause of mange of dogs, Sarcoptes scabiei var. canis In-vitro Diagnostic for Dog Contents of the Kit:
More informationEGFR (Phospho-Ser695)
Assay Biotechnology Company www.assaybiotech.com Tel: 1-877-883-7988 Fax: 1-877-610-9758 EGFR (Phospho-Ser695) Colorimetric Cell-Based ELISA Kit Catalog #: OKAG02090 Please read the provided manual entirely
More informationTaKaRa MiniBEST Viral RNA/DNA Extraction Kit Ver.5.0
Cat. # 9766 For Research Use TaKaRa MiniBEST Viral RNA/DNA Extraction Kit Ver.5.0 Product Manual Table of Contents I. Description... 3 II. Components... 3 III. Storage and shipping... 3 IV. Preparation
More informationDNA Purification for Case Transgene Pronuclear Injection Updated 2/26/08 RM
DNA Purification for Case Transgene Pronuclear Injection Updated 2/26/08 RM Materials: Stratagene StrataPrep DNA Gel Extraction Kit (Catalog #400766 or #400768) http://www.stratagene.com/products/displayproduct.aspx?pid=448
More informationMicroarray protocol Emmanuela Marchi PhD Dept. Pharmacology UFIR - Comparative nutritional systems biology Focus Team Post-doc emanuela.marchi@unifi.it 16 April 2009 When citing this SOP you should acknowledge
More informationMasterPure Plant RNA Purification Kit
MasterPure Plant RNA Purification Kit Cat. Nos. MPR09010 and MPR09100 Connect with Epicentre on our blog (epicentral.blogspot.com), Facebook (facebook.com/epicentrebio), and Twitter (@EpicentreBio). www.epicentre.com
More informationTechnical Note Detection of post-immunoprecipitation proteins by Western blot using the Quick Western Kit IRDye 680RD
Technical Note Detection of post-immunoprecipitation proteins by Western blot using the Quick Western Kit IRDye 680RD Developed for: Aerius, Odyssey Classic, Odyssey CLx and Odyssey Sa Imaging Systems
More informationMultiphor II Electrophoresis System
data file Electrophoresis Multiphor II Electrophoresis System One system for 1-D or 2-D protein electrophoresis or DNA fragment analysis Introduction Multiphor II Electrophoresis System and accessories
More informationDNA Extraction Kit INSTRUCTION MANUAL. Catalog # Revision B. For In Vitro Use Only
DNA Extraction Kit INSTRUCTION MANUAL Catalog #200600 Revision B For In Vitro Use Only 200600-12 LIMITED PRODUCT WARRANTY This warranty limits our liability to replacement of this product. No other warranties
More informationQS S Assist KINASE_MSA Kit
QS S Assist KINASE_MSA Kit Description KINASE MSA Kit is designed for use in pharmacological assays for KINASE based on Off-chip mobility shift assay (MSA). This kit includes Assay Buffer, Termination
More informationPlease read all the information in booklet before using the unit
100 Please read all the information in booklet before using the unit Bioneer Corporation 8-11,Munpyeongseo-ro, Daedeok-gu, Daejeon 34302, Republic of Korea Tel: +82-42-930-8777 Fax: +82-42-930-8688 Email:
More informationInstruction Manual for DNA-Microarrays
Instruction Manual for DNA-Microarrays 3-D Amino Surface Last update: 2004-12 page 1 Table of contents I. Introduction...3 II. Product description...4 A. Greiner Bio-One HTA Slides...4 B: SCIENION Buffer
More informationProductInformation. Genomic DNA Isolation Kit. Product No. GDI-3 Technical Bulletin No. MB-275 May 2000 TECHNICAL BULLETIN
Genomic DNA Isolation Kit Product No. GDI-3 Technical Bulletin No. MB-275 May 2000 TECHNICAL BULLETIN ProductInformation Product Description Sigma s Genomic DNA Isolation Kit isolates genomic DNA from
More informationNuclear Condensation Assay Kit Green Fluorescence
ab139479 Nuclear Condensation Assay Kit Green Fluorescence Instructions for Use Designed to assay chromatin condensation in live cells using an intercalating dye which is excitable with a standard 488nm
More informationFor Research Use Only Ver
INSTRUCTION MANUAL ZR-Duet DNA/RNA MiniPrep Plus Catalog No. D7003 Highlights Efficient isolation and separation of DNA and RNA from any cells, tissue, blood, and biological fluids. High quality DNA and
More informationab Plasmin Activity Assay Kit (Fluorometric)
ab204728 Plasmin Activity Assay Kit (Fluorometric) Instructions for Use For rapid, sensitive and accurate detection of Plasmin activity. This product is for research use only and is not intended for diagnostic
More informationqpcr Kit, DNA-free Product components 100 rxn 250 rxn Product description
qpcr Kit, DNA-free For the PCR detection and identification of bacterial and fungal DNA using custom primers Product code A8514 Product components 100 rxn 250 rxn A 2.5x mastermix (3 mm MgCl 2 final concentration)
More informationFor Research Use Only Ver
INSTRUCTION MANUAL Quick-RNA Whole Blood Catalog Nos. R1201 Highlights Purify high-quality total RNA (including small/micro RNAs) from whole and partitioned blood samples. Compatible with commonly used
More informationab Factor IXa Activity Assay Kit (Fluorometric)
ab204727 Factor IXa Activity Assay Kit (Fluorometric) Instructions for Use For rapid, sensitive and accurate detection of Factor IXa activity. This product is for research use only and is not intended
More informationCloning small RNAs for Solexa Sequencing version 2.0 by Nelson Lau Page 1 of 5 (Modified from Solexa sequencing protocol from Bartel lab)
Cloning small RNAs for Solexa Sequencing version 2.0 by Nelson Lau 09162008 Page 1 of 5 General Cloning Protocol: Gel-purification 1. Pour 1mm thick, urea denaturing 10% or 15% polyacrylamide gels, with
More informationProduct Information. Before you begin. Component A 1 vial of 30 ul vial of 300 ul each Glycerol. Tris
Glowing Products for Science Mix-n-Stain Antibody Labeling Kits Size: 1 labeling per kit Storage: -20 o C Stability: Stable for at least 1 year from date of receipt when stored as recommended. Components:
More informationab83355 ATP Assay Kit (Colorimetric/ Fluorometric)
Version 18 Last updated 2 January 2018 ab83355 ATP Assay Kit (Colorimetric/ Fluorometric) For the rapid, sensitive and accurate measurement of ATP in a variety of samples. This product is for research
More informationPositively Charged Membrane
BIOBOND NYLON MEMBRANES ProductInformation Technical Bulletin No. MB-570 June 1999 Size Quantity Positively Charged Membrane Neutral Membrane 30 cm x 3.5 m 1 roll N4781 N1031 30 cm x 12 m 1 roll N4906
More informationDetermination of Isoelectric Point (pi) By Whole-Column Detection cief
Determination of Isoelectric Point (pi) By Whole-Column Detection cief Tiemin Huang and Jiaqi Wu CONVERGENT BIOSCIENCE Determination of Isoelectric Point (pi) by Whole Column Detection cief Definition
More informationQIAGEN Supplementary Protocol: Isolation of genomic DNA from tissue using the QIAGEN-tip Storage of tissue samples.
QIAGEN Supplementary Protocol: Isolation of genomic DNA from tissue using the QIAGEN-tip 2500 This protocol is designed for the rapid, easy, and non-toxic preparation of up to 2 mg genomic DNA from not
More informationTransIT-TKO Transfection Reagent
Quick Reference Protocol, MSDS and Certificate of Analysis available at mirusbio.com/2150 INTRODUCTION TransIT-TKO is a broad spectrum sirna transfection reagent that enables high efficiency sirna delivery
More informationTITLE: LIVE/DEAD VIABILITY FOR CLINICAL SAMPLES
Paul K. Wallace, Ph.D. Director Roswell Park Cancer Institute Elm & Carlton Streets Voice:(716) 845-8471 Buffalo, NY 14263 Fax:(716) 845-8806 FILE NAME FL-SRP-2090.00 Live/Dead Viability for Clinical Samples
More informationNUCLEON Blood Non Chloroform DNA Extraction Kits For Research Use Only. Instructions for Use
NUCLEON Blood Non Chloroform DNA Extraction Kits 44100 For Research Use Only Instructions for Use NUCLEON is a registered trademark of Gen-Probe Life Sciences Ltd. NUCLEON kits are manufactured by Gen-Probe
More informationPowerMax Soil DNA Isolation Kit
PowerMax Soil DNA Isolation Kit Catalog No. Quantity 12988-10 10 Preps Instruction Manual Inhibitor Removal Technology (IRT) is a registered trademark of MO BIO Laboratories, Inc. and is covered by the
More informationab Plasmin Inhibitor Screening Assay Kit (Fluorometric)
ab204729 Plasmin Inhibitor Screening Assay Kit (Fluorometric) Instructions for Use For rapid, sensitive and accurate screening of potential Plasmin inhibitors. This product is for research use only and
More informationA Simpler and Faster Version of Two-Dimensional Gel Electrophoresis Using Vertical, Mini SDS-PAGE Apparatus
Iran. J. Chem. Chem. Eng. Vol. 28, No. 2, 2009 A Simpler and Faster Version of Two-Dimensional Gel Electrophoresis Using Vertical, Mini SDS-PAGE Apparatus Yaser Dorri* + ; Biji T. Kurien; R. Hal Scofield
More informationStandard Operation Procedure (SOP) for Biobanking Sampling Procedure Manual Use
Standard Operation Procedure (SOP) for Biobanking Sampling Procedure Manual Use 1. Oragene TM DNA Purification Protocol for use with Self-Collection kit of human sliva samples, OG-500 1.1. Equipment and
More informationSERVALight EosUltra Western Blot Chemiluminescence HRP Substrate Kit
INSTRUCTION MANUAL SERVALight EosUltra Western Blot Chemiluminescence HRP Substrate Kit (Cat. No. 42586) SERVA Electrophoresis GmbH Carl-Benz-Str. 7 D-69115 Heidelberg Phone +49-6221-138400, Fax +49-6221-1384010
More information