quick question about insoluble his-tag purification... - (May/09/2006 )

Hey all...pretty-please does anyone know the answer to this?

I am purifiying a protein under native conditions (his-tag, Qiaexpressionist methods). I am having solubility issues upon elution. I am amending my buffers and repurifying today. One of the tips is to conduct the purification steps at room temperature; we use sonication during the lysis step (no access to a french press )

this is my question: do you also sonicate at room temp? that is usually peformed on ice to prevent over-heating of the samples; I am concerned that removing the ice from this step would lead to temperatures greater than RT.

does anyone know the answer? I would be forever grateful.....

A

always sonicate on ice.

thanks, that's what I thought

I appreciate the assistance