Chemical compatibilities – Bio-Rad Nuvia™ IMAC Resin User Manual

Page 9

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Nuvia IMAC Ni-Charged Resin 5

Chemical Compatibilities

The chemical characteristics of Nuvia IMAC resin are detailed in Table 2.

Table 2. Chemical Compatibilities for Nuvia IMAC Resins.

Reagent

Group

Reagent

Comments

Stability

Buffer

reagents

Tris, HEPES,

MOPS

Sodium or

potassium

phosphate

Used with

proteins more

stable in

nonphosphate

buffers

≤50 mM

secondary and

tertiary amines

50 mM sodium

or potassium

phosphate are

recommended

as starting

buffers

Chelating

reagents

EDTA, EGTA

Strips nickel ions

from the resin

≤0.1 mM

successfully

used to remove

trace metal

contaminants

>1 mM can

cause significant

reduction in

binding capacity

Sulfhydryl

reagents

β-Mercaptoethanol

DTT, TCEP

Reduces random

disulfide bonds

preventing

protein

aggregation

during purification

Transition metals

at the center of

IMAC resin (Ni

2+

)

are susceptible to

reduction

≤20 mM

≤10 mM DTT

and 20 mM

TCEP

Detergents

Nonionic

detergents (Triton,

Tween)

Zwitterionic

detergents

(CHAPS, CHAPSO)

Removes

background

proteins

and nucleic acids

Solubilizes

membrane

proteins

≤2%

≤1%

Denaturants Guanidine HCl

(GuHCl)

Urea

Solubilizes

proteins

≤6 M

≤8 M

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