GE HEALTHCARE AKTAprime plus, Краткое руководство по эксплуатации

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11-0027-48 AC, 2007-09 • p9
Ordering information
Product Quantity Code No.
HisTrap HP 5 × 1 ml 17-5247-01
100 × 1 ml
*
17-5247-05
HisTrap FF 5 × 1 ml 17-5319-01
100 × 1 ml
*
17-5319-02
HisTrap FF crude 5 × 1 ml 11-0004-58
100 × 1 ml
*
11-0004-59
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.
For unclarified samples using HisTrap FF crude, make
sure the sample has been lysed properly, e.g. using
thorough sonication and DNase treatment.
•
System clogged – Replace the column with a piece
of tubing. Check pressure. If backpressure > 0.3 MPa,
clean system according to manual.
No binding:
•
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 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
5 10 15 20 25 30 35 min
2.5
2.0
1.5
1.0
0.5
0
UV 280 nm
Programmed %B
Sample: Clarified homogenate of
E. coli
expressing histidine-tagged
protein
Column: HisTrap™ HP 1 ml
Binding buffer (port A1): 20 mM phosphate, 0.5 M NaCl, 20 mM
imidazole, pH 7.4
Elution buffer (port B): 20 mM phosphate, 0.5 M NaCl, 0.5 M
imidazole, pH 7.4
100
80
60
40
20
0
% B
AU
280 nm
*
Pack size available by special order