ACS BIOCHEMISTRY EXAM Questions and Answers Henderson -Hasselbach Equation Ans- pH = pKa + log ([A -] / [HA]) Salting Out (Purification) Ans- Changes soluble protein to solid precipit ate. Protein precipitates when the charges on the protein match the charges in the solution. Size-Exclusion Chromatography Ans- Separates sample based on size with smaller molecules eluting later. Ion-Exchange Chromatography Ans- Separates sample based on charge. CM attracts +, DEAE attracts -. May have repulsion effect on like charges. Salt or acid used to remove stuck proteins. Affinity Chromatography Ans- Attach a ligand that binds a protein to a bead. Elute with harsh chemic als or similar ligand. SDS-PAGE Ans- Uses SDS. Gel is made from cross -linked polyacrylamide. Separates based off of mass with smaller molecules moving faster. Visualized with Coomassie blue. DTT (dithiothreitol) Ans- Reduces disulfide bonds. Homolog s Ans- Shares 25% identity with another gene Orthologs Ans- Similar genes in different organisms Paralogs Ans- Similar "paired" genes in the same organism glycine Ans- gly, g alanine Ans- ala, a leucine Ans- leu, l isoleucine Ans- ile, i Proline Ans- Pro, P valine Ans- val, v Methionine Ans- Met, M Phenylalanine Ans- Phe, F tyrosine Ans- tyr, y
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ACS BIOCHEMISTRY EXAM Questions and Answers
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Henderson-Hasselbach Equation Ans- pH = pKa + log ([A-] / [HA]) Salting Out (Purification) Ans- Changes soluble protein to solid precipitate. Protein precipitates when the charges on the protein match the charges in the solution. Size-Exclusion Chromatography Ans- Separates sample based on size with smaller molecules eluting later. Ion-Exchange Chromatography Ans- Separates sample based on charge. CM attracts +, DEAE attracts -. May have repulsion effect on like charges. Salt or acid used to remove stuck proteins. Affinity Chromatography Ans- Attach a ligand that binds a protein to a bead. Elute with harsh chemicals or similar ligand. SDS-PAGE Ans- Uses SDS. Gel is made from cross-linked polyacrylamide. Separates based off of mass with smaller molecules moving faster. Visualized with Coomassie blue. DTT (dithiothreitol) Ans- Reduces disulfide bonds. Homologs Ans- Shares 25% identity with another gene Orthologs Ans- Similar genes in different organisms Paralogs Ans- Similar "paired" genes in the same organism glycine Ans- gly, g alanin
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ACS BIOCHEMISTRY EXAM Questions and Answers
