Recently, my professor has given me a task to learn how to operate the FPLC system. I plan to run protein samples either HIC or IMAC column. I have ran tiny spin columns in the past, so I know how to prep the buffers. I looked over the user manual for the device and the columns. The system will be operating on a software called Unicorn 5.31. I familiarized myself with the software by playing around with it. I scan over the index and learned the complexity of it. I learned that the purification process can be automated, but how do I begin trying to automated my own process?
I would recommend that you take instruction from someone with a long experience of using the equipment. If you can't find someone at your institution, contact GE Healthcare Life Sciences and ask about taking a training course. There isn't much you can do to hurt the FPLC, but it is easy to ruin the columns if you aren't careful, and they can be expensive. Also, there are important aspects to learn about optimizing the separation.
I would recommend that you take instruction from someone with a long experience of using the equipment. If you can't find someone at your institution, contact GE Healthcare Life Sciences and ask about taking a training course. There isn't much you can do to hurt the FPLC, but it is easy to ruin the columns if you aren't careful, and they can be expensive. Also, there are important aspects to learn about optimizing the separation.
Basically you need to make your own mistakes as in any learning process. Don’t use pre-pack columns at this step. Get several resins and empty columns, make your own protein standards and start “playing". Don’t forget to filter and degass everything and anything or you’ll be clogging the system. Fix your own experimental conditions (sample load, pressure, resins, buffers) until you get sharp peaks and your results are reproducible. When you get to this point you’ll feel pretty familiarized with the equipment and you can go and use the expensive pre-pack columns. Before you get to the HIC, make sure you can use ion exchange.
Ismaila,
Search for an online course on Youtube or contact the manufacturer.
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