Testing for cracks in the column is quite easy. Take whichever standard you happen to have (lisozyme, GST, etc) and do a run. If it comes out in a single peak the problem is with your prep. If it comes out throughout the run the problem is with your column.
Mario Sanches On Sun, Aug 28, 2011 at 5:35 PM, Nian Huang <huangn...@gmail.com> wrote: > Hi, David, > What is the common cause of knackered SEC column? Will equilibrizing a > buffer containing 150 mM NaCl directly into a 20% EtOH or vise versa cause > the problem. There was no problem just after packing the column. > > Nian > > > On Sun, Aug 28, 2011 at 9:24 AM, David Briggs <drdavidcbri...@gmail.com>wrote: > >> Following on from Roger's fine suggestions: >> >> 8. Your column is knackered. Can you see fine lines or cracks in the >> column? Good packing is v.important for SEC columns. >> >> HTH, >> >> Dave >> >> >> ============================ >> David C. Briggs PhD >> Father, Structural Biologist and Sceptic >> ============================ >> University of Manchester E-mail: >> david.c.bri...@manchester.ac.uk >> ============================ >> http://manchester.academia.edu/DavidBriggs (v.sensible) >> http://xtaldave.wordpress.com/ (sensible) >> http://xtaldave.posterous.com/ (less sensible) >> Twitter: @xtaldave >> Skype: DocDCB >> ============================ >> >> >> >> On 28 August 2011 10:25, Allan Pang <a.p...@qmul.ac.uk> wrote: >> > Hi there everyone, >> > >> > What does it mean when you have proteins eluting in almost the whole >> column >> > volume of S200? >> > >> > I ran a gel with fractions from 8ml to 20ml and saw band for my protein >> all >> > throughout. >> > >> > Judging peaks on chromatogram is not useful as it doesn't have any >> aromatic >> > rings. >> > >> > Cheers, >> > >> > Allan >> > >> > -- >> > Allan Pang >> > >> > PhD Student >> > >> > G35 Joseph Priestley Building >> > Queen Mary University of London >> > London >> > E1 4NS >> > >> > Phone number: 02078828480 >> > >> > > -- Mario Sanches Postdoctoral Fellow Samuel Lunenfeld Research Institute Mount Sinai Hospital 600 University Ave Toronto - Ontario Canada M5G 1X5 http://ca.linkedin.com/in/mariosanches
