AKTA purifier Prgramming - (Nov/21/2008 )

Hello Everyone
I am new user to AKTA purifier and wish to programme it as follow-
start concentration of B 45% should run for 100 ml volume
then Concentration of B should reach to 60% and run for 50 ml
then concentration of B should reach to 100% in next 50 ml (where protein will be eluted from column)

Can anybody please suggest me which type of gradient i should opt for on AKTA
I tried to use segmented gradient but machine got stucked at 45% of B and it never went above that concentration. I tried several times reprogramming it in vain.

It will great if somebody please give suggestions in this context

Thanks
rang

did you program multiple gradient steps?

which version of unicorn are you using (i have an old version).

I can't remember if the gradient setting up according to the volume or the time.

I think it is a time issue. I will check it tomorrow when I go to work.

So set 45% B 100ml ( for example, if your flow rate is 2ml/min, then you should do 45% B, 50min), etc...