Bio-Rad Bio-Gel P Polyacrylamide Gel - Manual (Page 6)

Table 2. Formulations for buffers and solutions provided in the
Native IMAC Kits.
Profinia Solution Supplied Concentrated Formulation Working 1x Formulation Volume Position #
As
Native IMAC 2x 600 mM KCl, 100 mM KH
2
PO
4
, 300 mM KCl, 50 mM KH
2
PO
4
,
lysis buffer 10 mM imidazole, pH 8.0 5 mM imidazole, pH 8.0 125 ml N/A
Native IMAC 2x 600 mM KCl, 100 mM KH
2
PO
4
, 300 mM KCl, 50 mM KH
2
PO
4
,
wash buffer 1 10 mM imidazole, pH 8.0 5 mM imidazole, pH 8.0 125 ml 1
Native IMAC 2x 600 mM KCl, 100 mM KH
2
PO
4
, 300 mM KCl, 50 mM KH
2
PO
4
,
wash buffer 2 20 mM imidazole, pH 8.0 10 mM imidazole, pH 8.0 100 ml 2
Native IMAC 2x 600 mM KCl, 100 mM KH
2
PO
4
, 300 mM KCl, 50 mM KH
2
PO
4
,
elution buffer 500 mM imidazole, pH 8.0 250 mM imidazole, pH 8.0 100 ml 3
Desalting buffer 5x 685 mM NaCl, 13.5 mM KCl, 137 mM NaCl, 2.7 mM KCl,
21.5 mM Na
2
HP0
4
, 4.3 mM Na
2
HP0
4
,
40.5 mM KH
2
HP0
4
, 8.1 mM KH
2
P0
4
, pH 7.4 200 ml 4
pH 7.0 (pH 7.4 upon dilution)
Cleaning solution 1 2x 1,000 mM NaCl, 500 mM NaCl,
100 mM Tris, pH 8.0 50 mM Tris, pH 8.0 125 ml 5
Cleaning solution 2 4x 2,000 mM NaCl, 500 mM NaCl,
400 mM NaOAc, pH 4.5 100 mM NaOAc, pH 4.5 125 ml 6
Storage solution 2x 4% C
6
H
5
CH
2
OH 2% C
6
H
5
CH
2
OH
(benzyl alcohol) (benzyl alcohol) 200 ml 7
2.2 Instructions for Use
The buffers are provided as concentrates and are automatically
proportioned to a 1x formulation by the Profinia instrument when using the
preprogrammed methods. Each bottle is clearly labeled with a white strip
near the graduations, which provides the buffer name and the buffer
position number for insertion (1–7). The position number on the bottle
must match the position number on the instrument to properly run the
preprogrammed methods. Prior to inserting buffers into the instrument, the
following steps must be performed. Remove cap closures from buffer bottles
and rest closures on top of instrument, directly above appropriate buffer
positions. Rest a green buffer lid on the opening of each bottle and carefully
guide the tubing into the bottle. Place buffer bottle in appropriate numbered
position. Buffer lids help prevent buffers from being contaminated with
particulates and minimize evaporation during purification runs. Refer to
Figure 1 for a diagram of proper bottle installation and placement. If fewer
than 10 runs are used for a single kit, the remaining buffers can be capped
and stored at 4°C for up to 3 months.
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