Buffer Exchange & Protein Purification - (Jan/28/2008 )

Once and for all, I really need to get this 'basic knowledge' correct...

1. Can i know how important it is to do buffer exchange after protein purification that contains imidazole. I was told that Imidazole interfere with spectro reading, and thus it will not be accurate if you are using protein 280 to quantitate your protein. Is this true?

2. Secondly, how about other methods of quantification (bradford, BCA, Lowr and etc), will imidazole still in a way? Most company lists down the compatibility substance... do we still need to change buffer?

3. If my end point is to bring my protein for ELISA, is this still relevant?

Thank you in advance for the advice.


Haunted by Imidazole: spices

I had the same problem too.

imidazole dose interfere with the specto reading, i tested my protein before and after dialysis and got big different in

reading.

I personally prefer protein quantification in BCA or bradford methods, its much accurate. imidazole, DNA or RNA dont

interfere with the reaction.