AKTA Purifier
Contents
Section titled “Contents”1. Overview
Section titled “1. Overview”
The AKTA system is great for purifying proteins. We have many different columns that should suit your needs. Affinity columns, Q and SP, Gel Filtration columns for size exclusion and a set of metal columns (just cobalt (pink)). Feel free to pack your own columns and make great proteins/do good science.
2. General protocol
Section titled “2. General protocol”This protocol will change depending on the column you use and the protein you are trying to purify.
- Make your buffers A (low salt) and B (high salt) and filter sterilize them!
- Store the buffers at 4C.
- Run the pumpwash program for lines A1 and B1 to clear the pumps of ethanol or old buffer.
- Attached your column to the AKTA and equilibrate the column in your buffer A. (If your column is in ethanol, you must run a 10 column volumes of water through it first or else any salt buffer will crash and you will ruin the column.)
- Load your protein onto the column using a loop or a super loop. (collect the flowthrough? up to you)
- Wash the column with 10 column volumes of buffer A.
- Run a gradient from Buffer A to Buffer B.
- Collect fractions? up to you.
3. Available columns
Section titled “3. Available columns”Please look these columns up on GE Life Sciences website to find out information on pressure, flow and what the columns are used for. You can also look on the side of the deli case where all the column information is posted.
- Resource Q :17-1177-01
- HiTrap Heparin HP : 17-0407-03
- HiTrap SP XL : 17-5161-01
- HisTrap FF (5mL): 17-5255-01
- HisTrap FF (1mL):17-5319-01
- HiTrap Q XL : 17-5159-01
We also have several resins for packing your own columns.
- Talon Affinity Resin
- SuperDex 200
- Illustra Sephacryl S-1000
4. Maintenance
Section titled “4. Maintenance”- Manual
- For Preventative Maintenance and Service Calls, use the instructions below. Contract expires 9/01/2020