FPLC problem
Posted: Fri Jul 10, 2009 3:35 pm
OK I have a problem with AKTA FPLC system with Superdex 75 GL 10/300 column since this morning. I started equilibrating the column (25mM Bicine Buffer, 100mM NaCl, 1mM CHAPS, pH=9,5) at 0,5ml/min. Than I tried to put 0,75ml/min which normally works but there was activation of the pressure alarm (max 1,8 MPa) so I restarted the equilibration at 0,5ml/min and noticed UV baseline oscillation with unnaturally high value around 15mAU?? So I did Autozero UV. Even without flow UV values oscillates and started to grow, actually 0,1681, still growing). What could be the problem? Thanks for helping me.