Protein Expression and Purification Series - Bio-Rad

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CHAPTER 8 BIOLOGIC DUOFLOW PROTOCOL Protein Expression and Purification Series Purification Workflow for BioLogic DuoFlow System Preparing SDS-PAGE Samples of Desalted GST-DHFR-His Fractions Prepare SDS-PAGE samples. Add 25 µl of each fraction to 25 µl Laemmli buffer in 2 ml screwcap tube. Mix well. Heat all samples at 95°C for five minutes. + 25 µl fraction Desalted A PAGE SDS-PAGE sample Desalted B PAGE Heat 2 min at 95°C + 25 µl Laemmli buffer Desalted C PAGE 198 Chapter 8: Purification Protocol for BioLogic DuoFlow System
Protein Expression and Purification Series Purification Workflow for BioLogic DuoFlow System Quantitation of Protein in Desalted Fractions Turn on spectrophotometer and set absorbance to 280 nm. Add 100 µl distilled water to clean UV compatible cuvette. Blank spectrophotometer with distilled water. Add 100 µl Desalted fractions A, B, and C each into its own clean UV compatible cuvette. Measure absorbance of each fraction at 280nm. Return fraction to 2 ml tube. Calculate concentration of GST-DHFR-His in each desalted fraction as described in instruction manual and record data. Chapter 8: Purification Protocol for BioLogic DuoFlow System 199 + 100 µl dH 2 O + 100 µl Desalted A, B, or C Clean UV cuvette Clean UV cuvette CHAPTER 8 BIOLOGIC DUOFLOW PROTOCOL
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Biotechnology Explorer TM Protein E
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Protein Expression and Purification
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CHAPTER 1 BACKGROUND Protein Expres
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CHAPTER 2 FOCUS QUESTIONS Protein E
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CHAPTER 3A ADVANCE PREP CENTRIFUGAT
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CHAPTER 3B ADVANCE PREP INSTRUMENTA
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CHAPTER 4 11 ml CULTURE PROTOCOL Pr
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CHAPTER 5 100 ml CULTURE PROTOCOL P
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CHAPTER 6 HANDPACKING COLUMNs Prote
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CHAPTER 7 BIOLOGIC LP SYSTEM PROTOC
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Page 214 and 215: APPENDIX B RESULTS ANALYSIS Protein
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Page 218 and 219: APPENDIX C RCF TO RPM CONVERSTION P
Page 220 and 221: APPENDIX D SETTING UP THE SMARTSPEC
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Page 224 and 225: APPENDIX E MINI-PROTEAN GEL BOX SET
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Page 230 and 231: APPENDIX G BIOMANUFACTURING Protein
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Page 234 and 235: APPENDIX H FOCUS QUESTIONS INSTRUCT
Page 242 and 243: APPENDIX I GLOSSARY Protein Express
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Page 246 and 247: APPENDIX J REFERENCES, LEGAL NOTES
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Life Science Group Bulletin 1665067
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