Protein Purification by precipitation
Posted 26 August 2010 - 10:09 PM
Posted 26 August 2010 - 10:20 PM
Posted 27 August 2010 - 12:21 AM
Actually we perfrom both reducing and non reducing page and these are not multimers. yes to some extent the intensity of some band decreases but we get number of bands and we are unable to get rid of them.
Hola Firstly I thoudht that the upper bands were multimer, but you use denaturing PAGE,and I think this is the clue. Probably if you run a well withouth reducer it would appears a hihger band of your protein and the contaminant. So you have the contamination specifically bounded by disulphure bridge to your protein, wich you broke in PAGE. So add a bit of reducer in the tangential filtration or in the ion exchage buffer. If you put it I would increase it. Good luck