915
edits
Changes
→FPLC Run: Added details about HPLC run
==FPLC Run==
#Fill trey tray with ice, or put PBS and Glutathione in ice bucket.#Turn computer on and login if necessary, username=saltielhplc; password=eatAdEv3#Open TC Navigator and login#Go to Apps -> Kickoff -> GST Purification#Enter number of samples you want to run#Enter sample names and click through to finish#Start the pump, by going toHands On, Start Pump#Turn on lamp if necessary by going to Detector -> Turn Lamp On within the Hands On Window
#Rinse the leads with water and place in PBS (A) and Glutathione (B)
#Prime the leads by first selecting 100% A on the Hands On Window and while the pump is running, loosen the black screw in the lower section (remove the panel). Attach a 50 mL syringe and pull through 5-10mL
#Switch the pump to 100% B (Glutathione solution) and prime those lines. Switch back to 100% A (PBS)
#Connect the GSTrap column and let equillibrate for a few minutes
#WIth the injection loop set to load, using a syringe inject ~5mL of lysate into the 5mL sample loop
#Turn the loop clockwise to inject to start the run
#After 30min, the fraction collecter will automatically start, make sure it has 2mL tubes without caps ready
==Post-FPLC==