Care and use manual – Waters Protein-Pak Hi Res IEX Columns User Manual
Page 4
4
[ CARE AND USE MANUAL ]
Protein-Pak Hi Res IEX Columns and Standards
Protein-Pak Hi Res CM Column
AU
0.040
0.050
0.060
0.070
0.080
0.090
0.030
0.020
0.010
0.000
8
Lysozyme
Ribonuclease A
Cytochrome C
10
12
14
16
18
20
22
24 min
Protein-Pak Hi Res SP Column
Lysozyme
Ribonuclease A
Cytochrome C
AU
0.040
0.050
0.060
0.070
0.080
0.030
0.020
0.010
0.000
8
10
12
14
16
18
20
22
24
26
28
30
32
34 min
Protein-Pak Hi Res Anion-Exchange Column and Standard
Sample Diluent: Lyophilized in 20 mM Tris Buffer, pH 8–9,
Reconstituted in 1ml 100% Milli-Q Water, vortex and
inspect for undisssolved solids.
Protein
Sigma P/N
Conc. mg/mL
Horse Myoglobin
M1882
0.5
Conalbumin from
Chicken Egg White
C0755
1.25
Soybean Trypsin Inhibitor
T9128
2
Column:
Protein Pak Hi Res Q, 4.6 x 100 mm
Inj Vol:
10.0 µL
Flow Rate:
0.6 mL/min
Mobile Phase A (Acid):
100 mM TRIS HCl (aq)
Mobile Phase B (Base):
100 mM TRIS (aq)
Mobile Phase C (Salt):
1000 mM NaCl (aq)
Mobile Phase D (Aqueous): Water
Sample Manager Wash:
25 mM TRIS, 150 mM NaCl, pH 8.3
Sample Manager Purge:
25 mM TRIS, 150 mM NaCl, pH 8.3
Seal Wash:
90/10 H
2
O/MeOH
Column Temp.:
30 °C
Sample Manager Temp.:
4 °C
Detection:
280 nm
Auto•Blend Plus Gradient:
Time
(min)
Flow
(mL/min)
pH
pH
Curve
Salt
(mM)
Salt
Curve
Initial
0.6
9.0
6.0
0.0
6.0
2.0
0.6
9.0
6.0
0.0
6.0
15.0
0.6
9.0
6.0
500.0
6.0
18.0
0.6
9.0
6.0
500.0
6.0
19.0
0.6
9.0
6.0
0.0
6.0
20.0
0.6
9.0
6.0
0.0
6.0
Actual Gradient (with no Auto•Blend capability):
Time
Flow
(mL/min)
%A
%B
%C
%D
Initial
0.6
3.3
16.7
0.0
80.0
2.0
0.6
3.3
16.7
0.0
80.0
15.0
0.6
3.3
16.7
50.0
30.0
18.0
0.6
3.3
16.7
50.0
30.0
19.0
0.6
3.3
16.7
0.0
80.0
20.0
0.6
3.3
16.7
0.0
80.0
Protein-Pak Hi Res Q Column
Myoglobin
Conalbumin*
Trypsin Inhibitor
*Contains Isoform A and B which could be resolved under various pH conditions)