High gravity crush beads ? - (Sep/19/2012 )
I am using HisPur Ni-NTA beads for purification of His-Tagged proteins - BATCH method
We changed the centrifuge machine - and I just randomly used 7000 rpm for 5 min to settle down the beads
Is it possible that the beads might have been crushed by high gravity ?
The suggested gravity is 700g while i used 5000 g
The yield of protein is also low this time compared to my previous run.
Any experience with this kind of problem ?
-@bhijit-
@bhijit on Wed Sep 19 07:23:45 2012 said:
I am using HisPur Ni-NTA beads for purification of His-Tagged proteins - BATCH method
We changed the centrifuge machine - and I just randomly used 7000 rpm for 5 min to settle down the beads
Is it possible that the beads might have been crushed by high gravity ?
yes. you probably crushed the beads. as you said, your yield was poor and you will not be able to effectively reuse them.
-mdfenko-