Removing fat from tissue preps?
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?
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