Team:Washington/Notebook/50mL purification

Supernatant Protein Purification, 50mL

 * 1) Inoculate 50mL culture of TB with ~750uL overnight culture
 * 2) Grow a 37c until OD600: 0.4
 * 3) Inoculate cells with IPTG so that the final concentration is 0.5mM (25uL of 1M IPTG for 50mL culture)
 * 4) Grow cultures until OD600: 4, or use time points if looking for comparison in protein in supernatant
 * 5) Transfer culture to 50mL Falcon Tube
 * 6) Centrifuge at 8000rpm for 20 min to pellet cells
 * 7) Pour supernatant into 60mL syringe with 0.45uM filter attached
 * 8) Apply filtered supernatant to Ni-column (see Ni-column Set up for directions)
 * 9) Collect flow through and re-apply to column
 * 10) Wash column with 12mL PBS
 * 11) Elute column into 15mL condenser tube with 12mL PBS with 100mM imidazole
 * 12) Spin elutant in condenser tube until desired volume (~500uL) this is you purified protein.