Hello Everybody Hope every research is going well. I would like to ask you 2 doubts
1. Regarding Protein dialyis: I want my purified protein in 1xPBS. So after purify the protein,15ml I dialysed the protein against MQ water,5000ml for 1 hour at room temperature then I dialysed the protein in 1x PBS,5000ml at kept at 4 degree for overnight. The next day morning I collected the samples. I changed the dialysed buffer only once, If I do this way the purified protein is free of imidazole and the protein properly soluble in 1xPBS??? I did this way because I dont want use much PBS ? 2. The recombinant protein purification: The protein , which I am purifying most of the protein bind to Ni-NTA though the purification process was finished. I could say 70% protein I am getting in elution fractions remaining protein still stick to the beads. I wanted to reuse the columns for many times. So Could anyone please suggest any buffer, which can wash and elute the protein from the bead???? So I can reuse the column. If anyone answer me for my questions that would great help to me from them. Thank you very much in advance Cheers Sudheer _______________________________________________ Methods mailing list [email protected] http://www.bio.net/biomail/listinfo/methods
