Term
| What part of a fractionating protein colum is generally some kind of polymer (often a polysaccharide) |
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Definition
| What part of a fractionating protein colum is generally some kind of polymer (often a polysaccharide) |
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Term
| Chromatographric separations can separate proteins based on what things? |
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Definition
| Chromatographric separations can separate proteins based on what things? |
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Term
| In ion-exchange chromatography a positively charged matrix is called a _______ exchanger |
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Definition
| In ion-exchange chromatography a positively charged matrix is called a _______ exchanger |
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Term
| In ion-exchange chromatography a negatively charged matrix is called a _______ exchanger |
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Definition
| In ion-exchange chromatography a negatively charged matrix is called a _______ exchanger |
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Term
| You are separating proteins using ion-exchange chromatography, and you are using a cation exchanger. Put these proteins in order of elution: Protein A (net positive charge), Protein B (large net negative charge), Protein C (large net positive charge), Protein D (net negative charge) |
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Definition
| Protein B, Protein D, Protein A, Protein C |
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Term
| What can be done to the pH in order to increase the cationic character of a protein? |
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Definition
| What can be done to the pH in order to increase the cationic character of a protein? |
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Term
| At a pH abover a protein's pI, is the net charge of the protein positive or negative? |
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Definition
| At a pH abover a protein's pI, is the net charge of the protein positive or negative? |
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Term
| In ion exchange chromatography proteins are usually eluted from the column by a solution with decreasing salt concentration. True or False? |
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Definition
| In ion exchange chromatography proteins are usually eluted from the column by a solution with decreasing salt concentration. True or False? |
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Term
| What kind of chromatography seperates proteins by size? |
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Definition
| What kind of chromatography seperates proteins by size? |
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Term
| In size-exclusion chromatography which proteins elute first? Why? |
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Definition
| In size-exclusion chromatography which proteins elute first? Why? |
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Term
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Definition
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Term
| With increased salt concentration the protein-protein interactions are stronger than the solvent-protein interactions. True or False? |
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Definition
| With increased salt concentration the protein-protein interactions are stronger than the solvent-protein interactions. True or False? |
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Term
| What kind of polymer is used in size-exculsion chromatography? |
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Definition
| What kind of polymer is used in size-exculsion chromatography? |
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Term
| Affinity chromatography separates proteins based upon what? |
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Definition
| Affinity chromatography separates proteins based upon what? |
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Term
| You are separating proteins using affinity chromatography. You have three proteins (A, B, C), that each bind with a correpsonding ligand (a, b, c). If you eluted the column with a solution of ligand a, then ligand c, and finally ligand b, what would the order of elution for the proteins be? |
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Definition
| Protein A, Protein C, Protein B |
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Term
| What specific kinds of protein binding properties can be used to separate proteins using affinity chromatography |
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Definition
| ATP binding property, interaction with a specific DNA sequence, interation with another protein, interaction with ice |
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Term
| What is covalently coupled to the matrix in affinity chromatography? |
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Definition
| The selected ligand (which interacts with the protein). |
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Term
| What kinds of solutions can be used for elution in affinity chromatography? |
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Definition
| Salt solutions or ligand solutions (the ligand binds to the protein to release it from the resign bound ligand) |
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Term
| Addition of short amino acid sequences (tags) to the beginning or end of a target protein can facilitate what kind of purification? |
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Definition
| Affinity based purification |
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Term
| Adding six histidine residues to the beginning or end of a protein will allow it to bind tightly to what kind of matrix? |
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Definition
| One with Ni2+ ions attached |
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Term
| What does PAGE stand for? What does it do? |
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Definition
| Polyacrylamide gel electrophoresis. It is a method to determine the purity and molecular weigh of a protein |
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Term
| PAGE separates proteins based upon what? How is this done? |
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Definition
| Proteins are separated by their charge-to-mass ratio. The proteins move through a 'molecular sieve' of polyacrylamide facilitated by an electric field. They travel at a rate that depends directly on charge and inversely on size and shape |
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Term
| In page which proteins move faster, positively charged or negatively charged? |
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Definition
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Term
| What kind of dye is often used to visualize proteins after electrophoresis? |
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Definition
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Term
| Why is SDS (sodium dodecyl sulfate) added to the protein sample and gel buffers in PAGE? |
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Definition
| So that all proteins are denatured and posses a negative charge (all the proteins have essentially the same charge to mass ratio) |
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Term
| In the presence of SDS what does the rate of movement of proteins in an electric field depend on? |
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Definition
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Term
| Treatment of protein samples with beta-mercaptoethanol and heat ensures what? |
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Definition
| Complete separation of polypeptide chains (beta-mercaptoethanol reduces the disulfide bonds) |
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