255 - Frederiksen

More documents

Recommendations

Info

EDVO-Kit #<strong>255</strong> xxxPurification & Size Determination of gfp & bfpTable of ContentsPageExperiment Components 3Experiment Requirements 3Background Information 5Experiment ProceduresExperiment Overview 7Packing and Equilibrating the Column 8Partial Purification of gfp and bfp Proteins 9Sample Preparation for Denaturing SDS-gel Electrophoresis 10Electrophoresis of Proteins 11Staining the Gel 14Determination of Molecular Weights 16Study Questions 18Instructor's Guidelines 19Pre-Lab Preparations 20Electrophoresis Hints and Help 22Idealized Schematic of Results 23Study Questions and Answers 24Material Safety Data Sheets 25Protein Plus, FluoroGreen and FluoroBlue are trademarks of EDVOTEK, Inc.InstaStain, EDVOTEK, and The Biotechnology Education Company are registered trademarksof EDVOTEK, Inc.EVT 005077KEDVOTEK, The Biotechnology Education Company, and InstaStain are registeredtrademarks of EDVOTEK, Inc.. Ready-to-Load and UltraSpec-Agarose are trademarksof EDVOTEK, Inc.EDVOTEK - The Biotechnology Education Company® • 1-800-EDVOTEK • www.edvotek.com
Purification & Size Determination of gfp & bfpEDVO-Kit #<strong>255</strong>Components & RequirementsExperiment ComponentsThere is enough of each samplefor six (6) groups sharing threepolyacrylamide gels.Store componentsA-D, F, G in thefreezer.Component StorageA Cell Extract containing green protein (gfp) FreezerB Cell Extract containing blue protein (bfp) FreezerC Column Elution Buffer (10x) FreezerD Protein Molecular Weight Standards FreezerE Dry Matrix Room temperatureF Protein Denaturing Solution FreezerG 50% Glycerol Solution Freezer• Tris Glycine SDS Electrophoresis Buffer Room temperature• Protein InstaStain® sheets Room temperature• Protein Plus Stain Room temperature• Chromatography Columns Room temperatureNone of the components have been prepared from humansources.RequirementsAll components areintended for educationalresearch only. They are notto be used for diagnostic ordrug purposes, nor administeredto or consumed byhumans or animals.THIS EXPERIMENT DOESNOT CONTAIN HUMANDNA. None of the experimentcomponents are derivedfrom human sources.• Vertical Gel Electrophoresis Apparatus (optional)• D.C. Power Source (optional)• Automatic Micropipet (5 -50µl) and Tips(Cat. #638 Fine Tip Micropipet Tips recommended)• Balance• Ice Buckets and Ice• Long U.V. lamps (Cat. #969)• Ring stands and column clamps• 1ml pipets and pipet pumps• Microtest tubes• Polyacrylamide gels (3)*• Glacial acetic acid• Methanol• Distilled water*Polyacrylamide gels are not included for the electrophoresis part of thisexperiment. The experiment is designed for two student groups to share a gel.A total of 3 polyacrylamide gels (Precast polyacrylamide gels, Cat. #651) areEDVOTEK - The Biotechnology Education Company®1-800-EDVOTEK • www.edvotek.comFAX: (301) 340-0582 • email: info@edvotek.comEVT 005077K
Page 1: The Biotechnology Education Company
Page 5 and 6: Purification & Size Determination o
Page 8 and 9: EDVO-Kit #255Purification & Size De
Page 10 and 11: 10EDVO-Kit #255Purification & Size
Page 20: 26EDVO-Kit #255Material Safety Data

255 - Frederiksen

Create successful ePaper yourself

Delete template?

Save as template?