Protein Expression and Purification Series - Bio-Rad

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APPENDIX D SETTING UP THE SMARTSPEC PLUS Protein Expression and Purification Series 9. When the run has completed, the following will be displayed: Press for report =continue 10. Press Print to print your data (if you have paper) or press Cancel if you hand-recorded your data. Ready to read absorbance =Options 11. Remove the cuvette containing your GST-DHFR-His sample and NADPH in 1x PBS from the chamber. Add the DHF into the cuvette, quickly mix, and immediately place the cuvette into the chamber. Close the sample compartment door and press Read Sample. Note: If you do not have printer paper in your spectrophotometer, manually record each data point for all time points as they appear on the screen display. Display Absorbance: 150 A340=0.623 75 12. When the run has completed, the following will be displayed: Press for report =continue Example data for the 75 second time point 13. Press Print to print your data (if you have paper) or press Cancel if you hand-recorded your data. 14. Remove the cuvette from the instrument and dispose of the cuvette and the sample properly. 222 Appendix D: SmartSpec Plus Setup
Protein Expression and Purification Series Appendix E : Setting up a Mini-PROTEAN Tetra Gel Box using Mini-PROTEAN TGX Gels Mini-PROTEAN TGX Gel Set Up 1. Remove comb: Position both thumbs on the ridges of the comb. Remove the comb by pushing upward in one smooth continuous motion (Figure 1). 2. Remove tape: Pull the green tape gently to remove from the bottom of the cassette (Figure 1). 3. Rinse wells: Use a syringe wash bottle or a disposable transfer pipette to rinse the wells with running buffer. Straighten the sides of the wells, if necessary. Figure 1. Mini-PROTEAN TGX gel. Assemble the Mini-PROTEAN Tetra Cell Note: When running two gels only, use the Electrode Assembly (the one with the banana plugs), NOT the Companion Running Module (the one without the banana plugs). When running four gels, both the Electrode Assembly and the Companion Running Module must be used, for a total of four gels (two gels per assembly). 1. Set the clamping frame to the open position on a clean flat surface (see Figure 2A). 2. Place the first gel sandwich or gel cassette (with the short plate facing inward) onto the gel supports. Gel supports are molded into the bottom of the clamping frame assembly; there are two supports in each side of the assembly. Note that the gel will now rest at a 30° angle, tilting away from the center of the clamping frame. Appendix E: Gel Box Setup 223 APPENDIX E MINI-PROTEAN GEL BOX SET UP
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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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CHAPTER 8 BIOLOGIC DUOFLOW PROTOCOL
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Page 172 and 173: CHAPTER 8 BIOLOGIC DUOFLOW PROTOCOL
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Page 210 and 211: APPENDIX A TIPS AND HELPFUL HINTS P
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Page 214 and 215: APPENDIX B RESULTS ANALYSIS Protein
Page 216 and 217: APPENDIX B RESULTS ANALYSIS Protein
Page 218 and 219: APPENDIX C RCF TO RPM CONVERSTION P
Page 220 and 221: APPENDIX D SETTING UP THE SMARTSPEC
Page 224 and 225: APPENDIX E MINI-PROTEAN GEL BOX SET
Page 226 and 227: APPENDIX F BIOLOGIC LP SYSTEM SETUP
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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
Page 250: Life Science Group Bulletin 1665067
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Protein Expression and Purification Series - Bio-Rad

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