Fast purification of protein from DNA
Achim Recktenwald
achim at ibex.ca
Fri Jun 14 14:43:15 EST 1996
nobody at nowhere wrote:
>
> I have a problem of purification an enzyme from DNA inpurities. Do you know
> a fust and reliable method to solve the problem for less then 1ml volume of an
> enzyme solution without loss of protein's bioactivity?
> Thanks in advance,
> Dima Mozzherin
If the pI of your protein is high, use a tiny bit of anion-exchanger. The DNA should stick to
it, but not the protein. The pH of the incubation-pH should be at least > 7 and better ~1pH-unit
below the protein's pI.
If your protein's pI is low, use cation-exchanger. In this case your protein will stick to it,
but not the DNA. You can elute the protein by adding NaCl to ~500 - 1000mM.
Achim
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