same problem to me too

ch ch307689 at ohiou.edu
Wed Mar 6 23:49:15 EST 2002


I got the same problem. The truncated GST overwhelmed the protein pool and
therefore the amount of fusion protein after purification turned to be very
small. Did you have any idea to solve this problem?
I think the initial OD of the culture can be kept around 0.5 and the
induction can go up to 2 hrs but you need optimize it also. The temperature
can be kept either at RT or 28C according to the trouble shooting
instructions from Amershampharmacia. I did not find the optimized condition
yet. After changing the factors one at a time, I found shorter induction
time did help. Furthermore, you can add DTT at final conc. of 5mM before
cell lysis to improve binding to the beads.



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