How many times can I use the amylose column?
The most important variable in determining the useful life of the amylose resin is the amount of time it is in contact with trace amounts of amylase present in the crude extract (see 3.1). Under normal conditions (crude extract from 1 liter of cells grown in LB + 0.2% glucose, 15 ml column), the column loses 1–3% of its initial binding capacity each time it is used. If the yield of fusion protein under these conditions is 40 mg, this means that after 3 to 5 runs there would be a decrease in the yield. In practice, we often use a column 8 or 10 times before we notice a significant drop in the yield.
Related Questions
- Much of my fusion protein flows through the amylose column. Is there anything I can do to improve my fusions affinity for the amylose column?
- Much of my fusion protein flows through the amylose column. Is there anything I can do to improve my fusion’s affinity for the amylose column?
- How many times can I use the amylose column?
