isolating nuclei from fat tissue

[Keith Hoek -BIOCHEM]

mman at biosci.cbs.umn.edu (Michael Man) writes:
You could try a method used for obtaining nuclei from adipose tissue (guinea
pig) detailed in Enan and Matsumara, 1995, Biochemical Pharmacology, Vol 49
No 2, pp 249-261.

good luck


>hi, 
>	Does anybody have experience on isolating nulcei from fat tissue?
>I am currently struggling with this problem.  Fat, as a tough connective 
>tissue, is resistant to glass bounce homogenization.  I can only get very 
>tiny amount of nuclei from 20 mice!  It's almost depressing to look what 
>I got.  
>	People in my lab suggested to use collagenase to digest fat 1st.  But 
>I am concerned about the o/n incubation with collagenase may introduce 
>artifacts to my experiment (DNase 1 hypersensitive site analysis, 
>comparing fasted and fed mice).  The other option is to use Polytron at 
>low speed and hope that the nuclei will survive (I have serious doubt).
>	Your suggestion is greatly appreciated!

>michael
--
_____________________________________________________________________
Keith Hoek                                      hoek at biosci.uq.edu.au

mRNA transport          http://florey.biosci.uq.edu.au/~hoek/Pg1.html