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letters to nature NATURE |VOL 415 |17 JANUARY 2002 |www.nature.com 343
 

Summary: letters to nature
NATURE |VOL 415 |17 JANUARY 2002 |www.nature.com 343
Studies with H-2Kb
cells
H-2Kb
cells derived from the muscle of heterozygous H-2Kb
-ts-A58 transgenic mice17
were
differentiated18
and transfected with adenovirus containing control vector or dominant-
negative AMPK16
. Five days later, cells were treated with leptin (100 nM) for 2 h in HEPES-
buffered saline containing 5 mM glucose, and the AMPK activity and phosphorylation of
ACC were examined.
Measurement of catecholamine and AMP, ADP, ATP content
Soleus muscles were homogenized with 0.5 N perchloric acid and centrifuged. We
measured total catecholamine content in the supernatant by a radioenzymatic assay kit
(Amersham Pharmacia). AMP, ADP and ATP were determined by anion exchange
chromatography using a SMART system (Amersham Pharmacia)28
.

  

Source: Agard, David - Department of Biochemistry and Biophysics, University of California at San Francisco

 

Collections: Biotechnology; Biology and Medicine