Bio-Rad Profinity IMAC Resins User Manual

Page 7

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Resin Characteristics

The characteristics of Profinity IMAC resin are detailed in Table 1.

Table 1. Characteristics of Profinity IMAC Resin

* Dynamic binding capacity conditions (Q

10%

determination):

Column volume:

1 ml, (ID x H), 7 mm x 2.6 cm

Sample:

1.8 mg/ml 6xHis-tagged pure protein (32 kD)

Flow rate:

1 ml/min loading, and 2 ml/min washing and eluting

Loading buffer:

50 mM sodium phosphate, 300 mM NaCl, 5 mM imidazole, pH 8.0

Washing buffer:

50 mM sodium phosphate, 300 mM NaCl, 10 mM imidazole, pH 8.0

Elution buffer:

50 mM sodium phosphate, 300 mM NaCl, 250 mM imidazole, pH 8.0

Note: Dynamic binding capacity will vary from protein to protein.

** Maximum pressure test:

Profinity IMAC resin packed in a 1.1 x 30 cm Amicon column to a bed height of 20 cm with 20 mM sodium
phosphate buffer up to 43 psi (3 bar). Flow rates were increased stepwise by 200 cm/hr and held for 2 min
at each step. The pressure-flow curve for Profinity IMAC becomes nonlinear at pressures above 109 psi.

*** pH stability: Ligand density and protein BC are essentially unchanged after resin is treated with

pH 1–14 solutions/buffers for up to 200 hr.

Functional ligand

Iminodiacetic acid (IDA)

Base bead

UNOsphere base matrix

Form

50% suspension in 20% EtOH; comes precharged
with Ni

2+

or uncharged

Particle size

45–90 µm

Mean particle size

60 µm

Metal ion capacity

12–30 µmol Cu

2+

/ml Profinity IMAC resin

Dynamic binding capacity*

≥15 mg/ml resin

Recommended linear flow rate

<600 cm/hr at 25°C

Maximum operating pressure**

109 psi

pH stability, uncharged resin*** (up to 200 hr) 1–14

Chemical compatibility

See Table 2

Storage

4°C to ambient temperature

Shelf life in 20% EtOH

>1 year at ambient temperature

Operational temperature

4–40°C

Autoclaving conditions

0.1 M sodium acetate at 120°C for 30 min

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