Protein Expression and Purification Series - Bio-Rad

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APPENDIX E MINI-PROTEAN GEL BOX SET UP Protein Expression and Purification Series Fig. 2. Assembling the Mini-PROTEAN Tetra Cell Electrophoresis Module. 3. 4. Please use caution when placing the first gel, making sure that the clamping frame remains balanced and does not tip over. Now, place the second gel on the other side of the clamping frame, again by resting the gel onto the supports. At this point there will be two gels resting at an angle, one on either side of the clamping frame, tilting away from the center of the frame (see Figure 2B). Note: It is critical that gel cassettes are placed into the clamping frame with the short plate facing inward. Also, the clamping frame requires two gels to create a functioning assembly. If an odd number of gels (one or three) is being run, you must use the buffer dam (see Figure 2B). Using one hand, gently pull both gels toward each other, making sure that they rest firmly and squarely against the green gaskets that are built into the clamping frame. Make certain that the short plates sit just below the notch at the top of the green gasket. While gently squeezing the gel sandwiches or cassettes against the green gaskets with one hand (keeping constant pressure and both gels firmly held in place), slide the green arms of the clamping frame over the gels, locking them into place. Alternatively, you may choose to pick up the entire assembly with both hands, making sure that the gels do not shift, and simultaneously slide both arms of the clamping frame into place (see Figure 2C). 224 Appendix E: Gel Box Setup
Protein Expression and Purification Series 5. 6. Appendix E: Gel Box Setup The arms of the clamping frame push the short plates of each gel cassette up against the notch in the green gasket, creating a leak-proof seal (check again to make certain that the short plates sit just below the notch at the top of the green gasket). At this point, the sample wells can be washed-out with running buffer (Figure 2D). Note: If running more than two gels, repeat steps 1A–D with the Companion Running Module. Important Note: Do not attempt to lock the green arms of the clamping frame, without first ensuring that the gel cassettes are perfectly aligned and stabilized against the notches on the green gaskets of the module. To prevent the gels from shifting during the locking step, firmly and evenly grip them in place against the core of the module with one hand. Run the gel(s): Prepare, load and electrophorese the samples according to the instructions in Chapters 4 or 5, depending on whether you are performing the centrifugation process or chromatography instrumentation process for purification. At the completion of the run, disconnect the cell and remove the cassette. CAUTION: When running one or two gels only, DO NOT place the Companion Running Module in the tank. Doing so will cause excessive heat generation and prevent electrophoretic separation. Removing the Gel from the Cassette after the Gel has Been Run 1. Open the cassette: Align the arrow on the opening lever with the arrows marked on the cassette (see Figure 3). Insert the lever between the cassette plates at all four locations and apply downward pressure to break each seal. Do not twist the lever. Gently pull apart the two plates beginning from the top of the cassette. 2. Remove Gel: Gently remove the gel from the cassette. Figure 3. Opening the gel cassette. 225 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 174 and 175: 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
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Page 226 and 227: APPENDIX F BIOLOGIC LP SYSTEM SETUP
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Page 230 and 231: APPENDIX G BIOMANUFACTURING Protein
Page 232 and 233: APPENDIX G BIOMANUFACTURING Protein
Page 234 and 235: APPENDIX H FOCUS QUESTIONS INSTRUCT
Page 242 and 243: APPENDIX I GLOSSARY Protein Express
Page 244 and 245: APPENDIX I GLOSSARY Protein Express
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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