GE HEALTHCARE AKTAprime plus, Quick Reference Instructions

Page: 31 / 40

11-0027-48 AC, 2007-09 • p31
5
Typical result
Ordering information
Product Quantity Code No.
HiTrap Benzamidine FF 2 × 1 ml 17-5143-02
(high sub)
5 × 1 ml 17-5143-01
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:
•
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.
0 5 10 15 20 25 min
1.5
1.0
0.5
0
7
6
5
4
pH
UV 280 nm
pH
AU
280 nm
Sample: Human plasma
Column: HiTrap Benzamidine FF (high sub) 1 ml
Binding buffer (port A1): 50 mM Tris-HCl, 0.5 M NaCl, pH 7.4
Elution buffer (port B): 50 mM Glycine-HCl, pH 3.0
Human plasma without
trypsin-like serine proteases
Eluted trypsin-like
serine proteases