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High gravity crush beads ?

Ni-NTA Histag

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#1 @bhijit

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Posted 18 September 2012 - 11:23 PM

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 ?

#2 mdfenko

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Posted 19 September 2012 - 08:34 AM

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.
talent does what it can
genius does what it must
i do what i get paid to do





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