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[@bhijit]

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 ?

[mdfenko]

@bhijit, on 18 September 2012 - 11:23 PM, 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.