Problem with protein concentration
Posted 08 January 2004 - 02:30 AM
i am working in structural biology, thus i try to produce enough protein to obtain crystals ... as you know you have to reach a very high concentration to produce crystals ... the only problem is that i loose more than 90% of the protein when i concentrate it with spining concentration unit such as centricon. I can't concentrate the protein with ion exchange column because i need to obtain the protein in a very precise buffer
Posted 09 January 2004 - 02:22 AM
Posted 09 January 2004 - 02:26 AM
Posted 12 January 2004 - 06:04 AM
Either that or the membrane cut off maybe too high
Posted 15 January 2004 - 02:23 PM
Posted 03 February 2004 - 09:51 AM
Long ago I have encountered with same problem and now I know that good decision in such case is incubate centricons(after concentration step, and any systems that contain membranes) overnight.
Posted 04 February 2004 - 04:48 AM
or could I use a prolonged inverted centrifugation step to get back my concentrated protein?
Posted 04 February 2004 - 05:21 AM
Posted 04 February 2004 - 06:30 PM
so the best way to recover the concentrated sample is incubation followed by pipette out the retentate?
also, precipitation method is common to concentrate protein sample. mm...any people compared the centricon to precipitation method?
Posted 04 February 2004 - 11:33 PM
2. In such case (see topic), yes
Posted 23 February 2004 - 08:20 AM
Posted 01 April 2004 - 07:02 PM
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