GE HEALTHCARE AKTAprime plus, Quick Reference Instructions

Page: 17 / 40

11-0027-48 AC, 2007-09 • p17
Ordering information
Product Quantity Code No.
GSTrap FF 2 × 1 ml 17-5130-02
5 × 1 ml 17-5130-01
100 × 1 ml * 17-5130-05
GSTrap HP 5 × 1 ml 17-5281-01
100 × 1 ml
*
17-5281-05
GSTrap 4B 5 x 1 ml 28-4017-45
100 x 1 ml * 28-4017-46
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
* Pack size available by special order
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:
•
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
0.8
1.6
AU
280 nm
0
20
40
60
80
% B
0
0 12.5 25 min
––
UV 280 nm
–– Programmed %B
GST-
tagged
protein
Inject
Sample: Clarified homogenate of E. coli expressing
GST-tagged protein
Column: GSTrap™ FF 1 ml
Binding buffer (A1): 20 mM sodium phosphate, 0.15 M NaCl, pH 7.3
Elution buffer (B): 50 mM Tris-HCl, 10 mM reduced glutathione, pH 8.0