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The mitochondrial FAD-forming enzymes reside in two distinct monofunctional enzymes, which can be separated in soluble and membrane-enriched fractions.
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Ten μg of soluble proteins were separated in 10% polyacrylamide gels and electroblotted onto nitrocellulose membranes (Hybond-ECL, Amersham, GE Healthcare).
After lysis and centrifugation, the soluble fractions from leaf tissue were separated in 16% SDS polyacrylamide gel, and analyzed with immunoblots.
Cellular proteins were separated into soluble and insoluble fractions in 1% Triton X-100.
Membrane-associated proteins were separated from soluble proteins in two phases of Triton X-114 as described previously [ 43].
Bauxites develop either on rocks that are initially low in iron or on iron-rich rocks under circumstances in which organic matter or some other special factor renders iron sufficiently soluble to be separated from the aluminum minerals.
Peripherin is assembled from soluble precursors in a similar way [ 44], thus soluble peripherin precursors could be separated from the insoluble peripherin filaments, and the peripherin soluble/insoluble ratio determined.
Most other proteins remained soluble and could thus be separated.
Zr and Hf can also be separated by fractional crystallization of potassium hexafluorozirconate (K2ZrF6), which is less soluble in water than the analogous hafnium derivative.
The γ-isomer, which makes up 20 25 percent of this mixture, is more soluble than the other isomers in certain solvents and can be separated from them.
Precipitated proteins were separated from the soluble fraction by centrifugation at > 17,000 g for 15 minutes.
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