Removing fat from tissue preps?

A.F. Simpson AFS7 at le.ac.uk
Wed Jun 7 15:17:22 EST 2000


I have been preparing rat and mouse tissue for SDS-PAGE protein gels.

The basic method is:
Slice up 50-75mg tissue
Homogenise in 100ul/10mg buffer + protease inhibitors
Leave on ice
Spin out debris in refrigerated microcentrifuge
Use supernatant on gel

My problem is that I an getting a slimy layer at the top of the
eppendorf after the spin.  A second spin doesn't get rid of it and it's
almost impossible to leave behind in the tube - it sticks to the tip and
gets sucked in straight away.

I'm assuming it's fat.  As far as I can tell it's not causing any
problems with the SDS-PAGE, but it has a high 'yuk' factor and I'm
worried it might interfere with the protein assay.  Does anyone have any
suggestions for getting rid of it?

love
Anna






More information about the Methods mailing list