Protein Expression and Purification Series - Bio-Rad

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APPENDIX I GLOSSARY Protein Expression and Purification Series Lowry Protein Assay – An absorbance test used to measure the protein concentration in a sample. The assay relies on the reaction of protein with alkaline copper tartrate and Folin and the change in color of the sample. Luer-Lok – A standardized system of low-pressure fluid fittings used for making leak-free connections between a male-taper fitting and its mating female part on medical and laboratory instruments. Lyse – To break open a cell. Mixed-mode chromatography – A method of chromatography in which the resin used utilizes multiple chromatography techniques, such as a combination of anion and cation exchange properties, to resolve a mixture of biomolecules. Mobile phase – The liquid, solvent, buffer, or sample, that moves through the stationary phase or chromatography resin. Pellet – The insoluble components of a lysed cell that settle in the bottom of the centrifuge tube during centrifugation of the cell lysate. Polyhistidine tag – A series of histidine residues (usually 6) fused to a protein that aids protein purification because of its strong binding to nickel (IMAC) columns. Also known as a “His-tag.” Prepacked cartridge – A chromatography column that is prepacked with chromatography resin. The cartridge is capped, keeping the resin contained. The cartridge has a quick-connect fitting on its bottom and top allowing for easy connection to a syringe or chromatography pump or system. Protein assay – A test using the shift in absorbance of colorimetric dye to determine protein concentration. A spectrophotometer is required to perform the assay. Two popular protein assay methods are the Bradford Protein Assay and the Lowry Method. Which method to use is based on compatibility of the method with reagents in the sample buffer, as well as sensitivity. Resin bed – The settled, packed chromatography resin in a column. Sample – A mixture of biomolecules that is dissolved in a buffer and which is applied to a chromatography column. SDS-PAGE – Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis is a technique of separating a mixture of proteins based solely on its size with the use of a gel medium and an electric field. Size exclusion chromatography – Also known as gel filtration chromatography, desalting chromatography, and buffer exchange chromatography, size exclusion chromatography uses beads containing tiny holes, or pores, to separate a mixture of molecules based on its size. The larger molecules, which cannot fit through the holes, pass quickly around the beads whereas smaller molecules enter the holes and pass through the column more slowly. Sodium Dodecyl Sulfate – Also known as SDS, it is a strongly anionic detergent used to coat proteins and give them an overall negative charge. Soluble – The parts of the cell that are easily dissolved in water or buffer. Spin column – A small chromatography column that fits into a standard bench-top centrifuge and allows for quick purification of biomolecules. The column may be prepacked or be empty, allowing the user to choose the resin. 244 Appendix I: Glossary
Protein Expression and Purification Series Stationary phase – In chromatography, the stationary phase is the chromatography resin that is used to bind the molecule of interest or separate the sample mixture. Tertiary structure – 3-dimensional shape of a folded polypeptide, maintained by disulfide bonds, electrostatic interactions, and hydrophobic effects. Appendix I: Glossary 245 APPENDIX I GLOSSARY
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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 194 and 195: CHAPTER 8 BIOLOGIC DUOFLOW PROTOCOL
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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
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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