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An overview of three common buffer exchange techniques for glutathione s-transferase (gst) protein: dialysis, desalting column, and spin column methods. Each technique has its advantages and disadvantages, such as gentleness towards the protein, speed, cost, and ease of sample recovery. The document also includes a protocol for using spin columns for buffer exchange.
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Changing buffers for your GST.
1. Dialysis Tubing with pores that have size smaller than the protein but larger than the buffer contents. Advantages Gentle on the protein Inexpensive Can be done in cold room Not labor intensive Better for larger volumes Disadvantages Takes time. Couple days is best for complete buffer change Risk of losing sample while handling tubing Sample difficult to recover if lost through tubing or clips 2. Desalting Column Use size exclusion resin. Sephadex G25, or pre-packed PD-10 columns Advantages: Quick Disadvantages: incomplete desalting Not best for changing to a new buffer More expensive Dilutes protein 3. Spin Column Attach a micro column to an eppi tube Place sample in column and spin Buffer passes through membrane, protein is retained in column cup Advantages More complete than desalting if repeated serially Relatively fast Can also concentrate the protein at the same time Sample easily recovered if leakage occurs Disadvantages Limited volumes can be done at a time Expense Serial spins may require some time depending on cutoff size Risk of drying sample, risk of precipitation of protein Buffer inside beads GST passes around beads and elutes first without salts
We have the spin columns. These other options are possible if you find that your conditions need them for your project. We can order these things if you give us advance notice. Protocol for the spin columns