GE HEALTHCARE AKTAprime plus, Quick Reference Instructions

Page: 23 / 40

11-0027-48 AC, 2007-09 • p23
Ordering information
Product Quantity Code No.
HiTrap Protein G HP 2 × 1 ml 17-0404-03
5 × 1 ml 17-0404-01
HiTrap Protein A HP 2 × 1 ml 17-0402-03
5 × 1 ml 17-0402-01
HiTrap rProtein A FF 2 × 1 ml 17-5079-02
5 × 1 ml 17-5079-01
HiTrap MabSelect SuRe TM 5 x 1 ml 11-0034-93
HiTrap MabSelect
TM
5 x 1 ml 28-4082-53
HiTrap MabSelect Xtra
TM
5 x 1 ml 28-4082-58
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
0.4
0.8
1.0
AU
280 nm
3
4
5
6
7
pH
2
7.5 15.0
Inject
Mouse
IgG
1
Sample: Cell culture supernatant
containing mouse IgG1
Column: HiTrap Protein G HP 1 ml
Binding buffer (A1): 20 mM sodium
phosphate, pH 7.0
Elution buffer (B): 0.1 M glycine-HCl, pH 3.0
––
UV 280 nm
––
pH
min