Bio-Rad ChromLab™ Software - Manual (Page 162)

6 | Creating a Method
160 | NGC Chromatography Systems and ChromLab Software
Standard Method Templates
The workflow for standard method templates consists of the following phases:
 Equilibration
 Sample application
 Column wash
 Elution
 Column wash
 Re-equilibration for the next run
ChromLab includes the following eight standard method templates.
Table 6. Standard Method Templates
Method Template Explanation
Affinity Affinity chromatography is the separation of biomolecules
based on highly specific interactions. For example:
antibody/antigen or antibody/protein A, chelation
(polyhistidine-tag/nickel), enzyme/substrate
(glutathione-s-transferase/glutathione).
Anion Exchange Anion exchange chromatography uses a positively charged
column matrix to bind negatively charged protein molecules.
Proteins are eluted from the matrix using a gradient of
increasing ionic strength (typically NaCl). Select a buffer
system with a pH lower than the pI of the target protein to
enhance protein binding.
Cation Exchange Cation exchange chromatography uses a negatively charged
column matrix to bind positively charged protein molecules.
Proteins are eluted from the matrix using a gradient of
increasing ionic strength (typically NaCl). Pick a buffer system
with a pH higher than the pI of your target protein to enhance
protein binding.