Bio-Rad Profinity IMAC Resins - Manual (Page 21)

Table 6. Purification Method Suggestions
1 ml Cartridge 5 ml Cartridge
Step CV Flow Rate Flow Rate
Equilibrate 5 2 ml/min 10 ml/min
Lysate Load 5 to 10 2 ml/min* 10 ml/min*
Wash 1 6 2 ml/min 10 ml/min
Wash 2 6 2 ml/min 10 ml/min
Elute 5 2 ml/min 10 ml/min
Standard methods that are compatible with any
type of chromatography system are listed below. To
maximize binding capacity with large proteins
(>100 kD), for purification at 4°C, or for purifications
under denaturing conditions, the lysate load flow
rate* can be decreased (to 0.5 ml/min for the 1 ml
cartridge and 2 ml/min for the 5 ml). This will have
to be determined empirically for individual proteins.
1. Equilibrate the cartridge with 5 column volumes
(CV) of equilibration/wash buffer 1 at 2 ml/min.
2. Load the sample lysate at 2 ml/min.
3. Wash the cartridge with 6 CV of wash buffer
1 at 2 ml/min.
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