nulliespooner

Hello everyone I am a biology student and in out protein seperation course we did seperation on an ion exchanger first we washed the column 10 times with the buffer and then loaded our protein sample. after loading the sample we collected the volume eluted from the column into a "wash fraction". I have no idea why we did that... what could possibly be the reason to collect this fraction? it is not for positive protein (eluted at 0 mM NaCl)... so what then? why do we do it? and what is in there? Thanks!!!!

Hello everyone I am a biology student and in our protein seperation course we did gel filtration seperation on sephacryl s-200 column in the beginning we collected the void volume eluted from the column can someone please explain to me why we check the o.d of void volume? what could be there? also, we loaded different sample volumes - 1.5 ml and 0.5 ml what could happen if we overload or underload the column? if I load a very small protein sample, will it not get seperated well on the column? Thank you!!