Hi everyone,
I am purifying a his-tagged transcription factor using Ni-NTA agarose. After eluted from the column, 260/280 could reach to 1.9 or more. I am sure that the protein can be expressed. So my question is how to remove those nucleic acids? I have tried to use SP column but the protein was precipitated during dilution.
Run it through a anion exchange column; the DNA will bind almost at any pH or conductivity that is usable for protein bind/elute. You could try to find conditions in which your protein flows throug,, while the DNA binds.
You can use dnase and rnase in combination with nucleotide removal kits...
Abhishek Kumar this is true, but since it is a transcription factor, he/she (sorry, don't know if Yongliang Zhang is male or female) might want to use it with DNA in an assay or so.
Dnase or RNase should be inactivated after the reaction completion..@Achim Recktenwald..
Forgot to mention in my last one: an effective method that does not negatively affect the transcription factor.
Exactly, Abhishek Kumar , therefore I proposed anion exchange chromatography, as DNA is so strongly charged, it will definitely bind. If he/she can develop it to a flow-through method, it will be very easy to do.
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