Bio-Rad Profinity eXact™ Purification and Tag Cleavage Consumables - Manual (Page 10)

Table 4. Buffer composition.
Bind/wash buffer* 100 mM sodium phosphate, pH 7.2
Elution buffer* 100 mM sodium phosphate,
100 mM sodium fluoride, pH 7.2
Regeneration 100 mM phosphoric acid
Storage 0.02% sodium azide,
100 mM sodium phosphate, pH 7.2
* Add urea to the bind/wash buffer and elution buffer in order to purify proteins
under denaturing conditions.
Table 5. Triggering anions.
Anion Compound Fast Cleavage Slow Cleavage
F- NaF, KF 100 mM 5 mM
N
3
- NaN
3
10 mM 1 mM
NO
2
- NaNO
2
5 mM 1 mM
CO
2
- NaHCO
2
1000 mM 25 mM
Cl- NaCl, KCl >1000 mM 75 mM
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Section 5
Buffers and Methods
Bio-Scale Mini Profinity eXact cartridges 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, a
strong chaotrope (e.g. urea) is included in the
bind/wash buffer and elution buffer, allowing target
proteins to be purified in an unfolded or partially
folded state. The recommended buffer compositions
and triggering anions are provided in the following
tables. Note that some loss in binding capacity may
be observed when buffers used contain greater
than 2 M urea. Additionally, urea concentrations
greater than 4 M may result in decreased target
protein purity.
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