isolating nuclei from fat tissue
Keith Hoek -BIOCHEM
hoek at biosci.uq.oz.au
Thu Feb 22 20:55:35 EST 1996
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.
> 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
> 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!
Keith Hoek hoek at biosci.uq.edu.au
mRNA transport http://florey.biosci.uq.edu.au/~hoek/Pg1.html
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