BBEC Pressurize/Diffusion Separations
I wanted to attempt pressurized and diffusion separations of brain extract using the new movable wafers. I tried pressure cell separations of 1:20 BBEC (clarified brain extract) at 2psi for 40 min. The flow rates were better than the standards I was previously using – this could be due to the little bit of glycerol in the standards. Here is the gel:
There isn’t a clear separation here. Perhaps the pores are too large. It might help if I ran this on a lower percentage gel, which would enable better resolution for larger proteins.
Diffusion separations:
Again, no well defined separation using BBEC using w670. Standards could be deceiving because of the complexed proteins that make up the ladder. Also BBEC is lacking clean bands above 100kD. Separations with 670 using standards were at least >100kD. I will compare 670 histograms to previous membranes (ex. 319) that had stong BBEC separations.
A couple other things I should note: 1) there seems to be some variability again with the filtrate in the two different samples, and 2) we’re seeing slight reduction of small species in retentate again.