Bio-Rad Profinity IMAC Resins User Manual

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Section 8

Preparation of Clarified E. coli Lysate

Using Denaturing Conditions ..............................18

Materials ....................................................................18

Method ......................................................................19

Section 9

Medium-Pressure Column Purification

of Histidine-Tagged (His-Tagged) Proteins

Using Nondenaturing Conditions........................21

Materials ....................................................................21

Method ......................................................................22

Section 10

Medium-Pressure Column Purification —

Using an Imidazole Gradient to

Determine Optimal Purification of

His-Tagged Proteins ..............................................23

Materials ....................................................................23

Method ......................................................................24

Section 11

Medium-Pressure Column Purification of

His-Tagged Proteins Using

Denaturing Conditions..........................................27

Materials ....................................................................27

Method ......................................................................29

Section 12

Sample Preparation-Size Spin-Column

Purification of His-Tagged Proteins

Using Nondenaturing Conditions........................31

Materials ....................................................................31

Method ......................................................................32

Section 13

Sample Preparation-Size Spin-Column

Purification of His-Tagged Proteins

Using Denaturing Conditions ..............................34

Materials ....................................................................34

Method ......................................................................35

Section 14

Batch-Mode Purification of Histidine

Proteins — Gravity Flow .......................................37

Recommended Columns............................................37

Materials ....................................................................37

Resin Preparation .......................................................38

Method ......................................................................38

Section 15

Regenerating, Cleaning, Sanitizing,

and Storage ............................................................40

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