Bio-Rad Profinia™ Protein Purification Instrument User Manual
Page 128
Table 10.3. Formulations for buffers and solutions provided in the Profinia denaturing IMAC
kits.
Profinia Supplied
Concentrated
Working
1x
Supplied
Vol*,
Buffer
Solution
Formulation
Formulation
Vol, ml
ml
Position
Denaturing
1.4x
420 mM KCl,
6 M urea, 300 mM KCl,
176
250
N/A
IMAC lysis
70 mM KH
2
PO
4
,
50 mM KH
2
PO
4
,
buffer
7 mM imidazole,
5 mM imidazole,
pH 8.0
pH 8.0
Denaturing
1.4x
420 mM KCl,
6 M urea, 300 mM KCl,
176
250
1
IMAC
70 mM KH
2
PO
4
,
50 mM KH
2
PO
4
,
wash buffer 1
7 mM imidazole,
5 mM imidazole,
pH 8.0
pH 8.0
Denaturing
1.4x
420 mM KCl,
6 M urea, 300 mM KCl,
88
125
2
IMAC
70 mM KH
2
PO
4
,
50 mM KH
2
PO
4
,
wash buffer 2
14 mM imidazole,
10 mM imidazole,
pH 8.0
pH 8.0
Denaturing
1.4x
420 mM KCl,
6 M urea, 300 mM KCl,
88
125
3
IMAC
70 mM KH
2
PO
4
,
50 mM KH
2
PO
4
,
elution
350 mM imidazole,
250 mM imidazole,
buffer
pH 8.0
pH 8.0
Cleaning
2x
1,000 mM NaCl,
500 mM NaCl,
125
N/A
5
solution 1
100 mM Tris,
50 mM Tris, pH 8.0
pH 8.0
Cleaning
4x
2,000 mM NaCl,
500 mM NaCl,
125
N/A
6
solution 2
400 mM NaOAc,
100 mM NaOAc,
pH 4.5
pH 4.5
Storage 2x
4%
C
6
H
5
CH
2
OH 2%
C
6
H
5
CH
2
OH
200
N/A
7
solution
(benzyl alcohol)
(benzyl alcohol)
*After addition of urea.
121