What is the best elution solution for his-tag purification chromatography? glucose solution imidazole solution high salt solution a buffer with a pH of 7
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Identify the buffer solution that can be used for eluting a transcription factor bound to a DNA affinity column: a. high salt concentration b. high imidazole concentration c. mixture of free DNA nucleotides d. high sugar concentration
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Stock solutions provided are: 1 M HEPES buffer, pH 7.5 5 M NaCl 1 M imidazole 80% (v/v) glycerol Question: Calculate the volume of stock solutions required to make up the buffer solutions that will be used for protein purification. The solutions you need to prepare for purification are: i. Binding Solution A: Make up 50 mL of 50 mM HEPES buffer (pH 7.5), 300 mM NaCl, 5 mM imidazole, 5% (v/v) glycerol. ii. Wash Solution B: Make up 50 mL of 50 mM HEPES buffer (pH 7.5), 300 mM NaCl, 75 mM imidazole, 5% (v/v) glycerol. iii. Elution Solution C: Make up 10 mL of 50 mM HEPES buffer (pH 7.5), 300 mM NaCl, 500 mM imidazole, 5% (v/v) glycerol.
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The buffer that we use this week contains 10mM Tris, pH 8.0, 150mM NaCl. The elution buffer is the same as the breaking buffer, but it also contains 300mM imidazole. The instructor made 2L of elution buffer for all the students. The instructor had 500ml of 2M Tris (121.1 g/mole) (pH 8.0), 750ml of 2.5M NaCl (MW = 58.44 g/mole), and 350ml of 1.5M imidazole (68.08 g/mole). How much of each stock solution did they need? Show your calculations for credit.
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