/sci/ - Science & Math

Any biochemistfags here? I got a cytosolic protein that's being a bitch to purify, and I was wondering if anyone here could offer some advice. My bacterial lysate looks pretty cloudy even though it gets passed through the French Press twice--it's viscous as fuck. SDS after column chromatography shows that most of my protein is pretty much trapped in the cell pellet, so I guess there's a lot of inclusion bodies present.

Any ideas on how to prevent inclusion body formation / protein misfolding?

>cytosolic heterologous His-tagged protein in E. coli
>induction with 0.5 mM IPTG at 25 degrees celsius overnight
>lysis buffer is essentially saltwater (50 mM Tris pH 8.0 and a couple protease inhibitor tablets)
>i dont feel like dousing this shit with 8M urea either

>>9471587
>king of maths but also ignorant