Bio-Rad Profinia™ Protein Purification Instrument - 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.
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