To identify the reason for backpressure in the system follow the scheme on the right hand side. Also keep in mind these four most common sources of backpressure:
Tubings – Keep tubing as short as possible. Bigger inner diameters give less back pressure, but also increase the dead volume of the system. Always replace clogged and nicked tubings.
Column – Over time dirt can accumulate on the FPLC column, which can result in higher backpressure. Most FPLC columns can be efficiently cleaned with 1 M NaOH.
Viscous buffers – Viscous buffers can generate high back pressure, this can only be dealt with by decreasing the working flow rate
Viscous sample – Make sure to filter your sample before injection to reduce back pressure.
If you face constant clogging of your inline filter at the pump, due to dirty samples or unfiltered buffers,you can also replace the inline filter with an empty cartridge, which is also included in the accessory kit. The column can still be protected by an additional inline filter placed directly in front of the column (pore size 10 µm (A3379)).