Hi everyone,

I'm working with protein purification and planning to let my samples through a hydroxyapatite column. My samples contain glycerol and would like to know whether I can load my samples directly to the column or only after getting rid of the glycerol? Thanks you very much for the opinions. Really appreciate it.

Alys

i used glycerol with hppt column and experienced somewhat low binding to the column. but still working if the amount of the needed protein isnt so importnat. you can try a small scale batch method for example 5 ul (20%)of hppt to a small volume of your sample e.g. 50 ul let it to bind ,make several washes and run load the ohppt into gel electrophoresis, and check your target. you can pass your sample through the column and collect the flowthrough if there is no binding it will be benificial as you get the protein again and sure some contaminants are boud to the column. you may also try diluting your sample the glycerol is diluted and i think ohppt separations are not sensetive to sample volume.