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120 Redox Report bated for 15 min at room temperature with HEPES/

Summary:  120 Redox Report
bated for 15 min at room temperature with HEPES/
KOH buffer, pH 8.0, ATP-MgC12 (1 mM), amino
acids (each at 25 pM; all protein amino acids except
methionine) and additional redox agents and inhib-
itors as indicated. Labelling of synthesised proteins was
initiated by addition of 30 pCi of ^S-methionine
(>1000 Ci/mmol). Illumination, where present, was
provided by a 40 W fluorescent strip lamp at 50 cm
from the sample tubes. After 45 min, intact, labelled
chloroplasts were broken by brief osmotic lysis, thylako-
ids (photosynthetic membranes)and stroma (chloroplast
soluble phase) were separated by centrifugation, and
samples were dissolved in electrophoresis buffer. An
equal amount of protein was added to each lane of
the gel.
Mitochondria were isolated8t9from green leaves from
11-12 day-old peas and equal volumes (5 pl) of a
concentrated mitochondrial suspension were preincu-
bated for 15 min, on a shaker, at 25OC in a reaction


Source: Allen, John F. - School of Biological and Chemical Sciences, Queen Mary, University of London


Collections: Renewable Energy; Biology and Medicine