[Bioforum] Pressure limit AND Ni-NTA beads

Pepa Florez Pérez via bioforum%40net.bio.net (by snipeurope from hotmail.com)
Mon Aug 3 13:25:21 EST 2009


Hello everybody,
 
here it is my first question. I am using Ni-NTA agarose beads to purify a protein.
According to the manufacturer the limit pressure is around 2 psi (nothing), and I am running the column to 50 psi.
 
Can anyone please tell me about why does the pressure limit is important when dealing with affinity chomatography?
 
How can this (pressure limit too high) affect my purification?
 
Am I destroying the beads?
 
I am running the column (home-prepacked tricorn) on an akta-prime.
 
I would really appreciate any information since I couldn't find any info on the web.
 
Thank you very much,
 
Eva

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