GE HEALTHCARE AKTAprime plus, Краткое руководство по эксплуатации

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11-0027-48 AC, 2007-09 • p19
Ordering information
Product Quantity Code No.
StrepTrap HP 5 × 1 ml 28-9075-46
HiTrap Desalting 5 × 5 ml 17-1408-01
100 × 5 ml * 11-0003-29
HiPrep 26/10 Desalting 1
(53 ml)
17-5087-01
4
(53 ml)
17-5087-02
Superloop 10 ml 1 18-1113-83
Superloop 50 ml 1 18-1113-84
Superloop 150 ml 1 18-1023-85
Troubleshooting
High backpressure:
•
Column clogged – Clean the column according
to instructions. Make sure the sample has been
centrifuged and/or filtered through a 0.45 µm filter.
•
System clogged
– Replace the column with a piece
of tubing. Check pressure. If backpressure > 0.3 MPa,
clean system according to manual.
No binding:
•
Regenerate the column with 3 column volumes (CV)
water, 3 CV 0.5 M NaOH, 3 CV water, 5 CV binding
buffer before starting the run.
•
Check that the correct column is used.
•
Check that the inlet tubing from each buffer is
connected to the correct inlet port.
•
Check that the composition and pH of the buffers are
correct.
•
Check that the sample has been adjusted to the
binding buffer conditions.
•
Check that your sample contains target protein.
No elution:
•
Check that the inlet tubing from each buffer is
connected to the correct inlet port.
•
Check that the composition and pH of the buffers are
correct.
•
Use alternative elution conditions according to the
column instructions.
•
Check that your sample contains target protein.
5
Typical result
*
Pack size available by special order
0.5
1.0
1.5
2.0
AU
280 nm
20
40
60
80
100
%B
0 5 10 15 20 25 30 35
min
Sample: Clarified lysate of E. coli expressing Strep(II)-tagged protein.
Column: StrepTrap™ HP 1 ml
Binding buffer (port A1): 100 mM Tris-HCl, 150 mM NaCl, 1 mM EDTA, pH 8.0
Elution buffer (port B):
100 mM Tris-HCl, 150 mM NaCl, 1 mM EDTA,
2.5 mM desthiobiotin, pH 8.0
0 0