Bio-Rad Nuvia™ IMAC Resin - Manual (Page 11)

Bio-Scale Mini Nuvia IMAC Ni-Charged 7
Section 5
Buffers and Methods
IMAC methods can be run using either native or denaturing
purification protocols. Under native conditions, proteins are purified
using buffers that help retain the natural folded structure of the
target protein. Under denaturing conditions, strong chaotropic
agents (typically 6–8 M urea or 6 M guanidine) are added to the
buffers, allowing target proteins to be purified in their unfolded
states. The recommended buffer compositions and formulations
are provided in the following table.
Table 4. Suggested buffer composition.
NaCl, mM Na Phosphate,
mM
Imidazole,
mM
Urea, M
Native lysis/
wash buffer 1
300 50 5 NA
Native wash
buffer 2
300 50 25 NA
Native elution
buffer
300 50 500 NA
Denaturing lysis/
wash buffer 1
300 50 5 6
Denaturing wash
buffer 2
300 50 25 6
Denaturing
elution buffer
300 50 500 6
For all buffer formulations, adjust pH to 8.0 with KOH or H
3
PO
4
and filter through a 0.2 μm filter.
Native buffers can be stored up to 1 year at 4–22°C; denaturing buffers must be made fresh
and used within 7 days or frozen in aliquots at –20°C for later use.