Hi Sudheer, for the same protein, I'd re-use the column a several times. As impurities, contaminants and some protein will accumulate in the column, and some nickel gets washed off, the performance of the column will drop by time.
Then simply purge and recharge the column as I have described before (EDTA, water, nickel salt, water, buffer). As it's so horribly cheap, when you do it with homemade reagents (not only compared to the value of your samples), it might be reasonable, to perform "preventive maintenance" before each run. Before cleaning another protein on the same column, I'd do a more harsh cleaning (EDTA, water, trypsin, water, NaOH, water, recharge), as this might harm the resin a little. No need to buy expensive proprietary solutions advertised as "magic", however. HTH Wo (the kit hacker :) On Saturday, November 17, 2012 3:02:39 AM UTC+8, sudheer sangeetham wrote: > Hi > > > > Thank you for your suggestions. I have one more doubt, how long time I can > > use each column? Even in qiagen expressionist, they suggested to use 5 > > times of each column. After that shall I discard the resin or need to > > recharge it with recharging buffers? > > > > thanks in advance > > > > > > -- > > Sudheer Babu.S > > Research Fellow > > Institute of Biochemistry > > Biological Research Center > > Szeged,Hungary. _______________________________________________ Methods mailing list [email protected] http://www.bio.net/biomail/listinfo/methods
