I've just started a protein expression extraction. To check I was getting protein expressed, I took 1 ml of my culture, pelleted the cells and resuspended in 2x SDS-PAGE loading dye. For uniduced, I added 50 ul of dye, for the 3hr induction I added 800 ul of dye. I'm kind of following an NEB pMAL guide to do this.

I ran the samples on a pre-bought and still in date Bis-Tris 12 % gels for 1 hr at 180 V. I loaded 10 ul of sample into each well (half what NEB recommend). For my induced samples (lanes with + above) the lanes are thin with a slight smear. I know that smears can come with protein overloading, but i'm not sure why the samples are running thin like this. Any suggestions?

Note: I've done similar expression pilot experiments, and resuspended 1 ml of 3hr induction culture in 500 ul and loaded 20 ul of sample and it worked beautifully.