You can run a few µl of your sample on a native gel or agarose gel and
visualise co-purified DNA by ethidium bromide/gel red staining.
If your protein is stable at high salt concentrations (and a lot of DNA-binding
proteins are) you can use a high salt concentration (like 1 M) in the lysis
buff
I'd agree with Pascal as well, and can add that if you have a protein
for which you use an affinity tag, often high salt (>500mM) will strip
the DNA off your protein, and might even keep it stable.
Cheers -
Miles
On Aug 10, 2009, at 3:59 PM, Pascal Egea wrote:
Hi Neeraj,
An absorption sp
toxin-free.
--Chun
Competing Interests Statement
Accelagen is the maker of TurboNuclease.
_
From: CCP4 bulletin board [mailto:ccp...@jiscmail.ac.uk] On Behalf Of Pascal
Egea
Sent: Monday, August 10, 2009 4:00 PM
To: CCP4BB@JISCMAIL.AC.UK
Subject: Re: [ccp4bb] DNA bi
I had a simple question about DNA binding protein. Is there an
easy way to detect if your heterologously expressed protein is bound to
DNA post purification.
Yes. UV absorbance. DNA absorbs UV strongly, proteins do not. DNA absorbs
260 more thn 280, the opposite is true for proteins. I
Hi Neeraj,
An absorption spectra between 220 and 400 nm (for example) should show you
if there is DNA coming along with your protein. In theory A280 is about 1.7
times A260 for a pure protein sample. This is a rough estimate. If your peak
is shifted towards 260 instead of 280 then you can suspect t
Hi all,
I had a simple question about DNA binding protein. Is there an
easy way to detect if your heterologously expressed protein is bound to
DNA post purification. Also is there an easy way to strip the protein of
DNA without any damage done to the protein in doing so. I would
apprec
