CaYPD1 purification-3Pre Lab

CaYPD1 purification-3Pre Lab - 6 mL of buffer 5 followed by...

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Pre Lab CaYPD1 purification-3 Purpose: Purification of dry cell pellet of His6-CaYPD1 or CaYPD1-His6. Techniques: To produce slurry invert the column gently. Dispose of waste in the designated containers. Steps: Take Ni-NTA column wash with 20mL Buffer 1 in to a beaker, then with 7mL buffer 2 in to test tube. Label test tube. Seal the test tube and store in the cold room. Elute the protein with 4 mL Buffer 3, followed by 4mL buffer 4 and 4mL of Buffer 5. Use 3 separate to hold the flow-through. Label and seal and store them. Wash the resin with
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Unformatted text preview: 6 mL of buffer 5 followed by 10 mL dH2O. Collect the flow though in beaker 1. Then wash the resin with 30mL .5M NaOH collect the flow in beaker 2. Wash the resin with another 10mL dH2O. Lastly, wash the resin with 10mL 20% ethanol. Collect the flow in beaker 1. Waste in the beakers should be disposed in the designated containers. Load several mL 20% ethanol into column. Cap the column, invert it to generate slurry. Pour it in the designated container. Wash the column and lids and return them....
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This note was uploaded on 10/25/2011 for the course ALL 101 taught by Professor All during the Spring '11 term at Cameron University.

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