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The preparations were transferred in normal Mac Ewen solution.
The size-fractionated antigen preparations were transferred to Immobilon polyvinylidene fluoride transfer membranes (Millipore, Bedford, MA, USA) as described by Towbin et al. (16 ).
Prior to electrophoresis, microsomal preparations were transferred to two-fold buffer, containing 0.125 M Tris-HCl (pH 6.8), 4 % sodium dodecyl sulphate (SDS), 20 % glycerol, 200 mM DTT and 0.02 % bromophenol blue.
Preparations were transferred to a recording chamber and a double lumen cannula was placed into the descending aorta for retrograde perfusion with a Ringer solution containing in mM: NaCl (125), NaHCO3 (24), KCl (3), CaCl2 (2.5), MgSO4 (1.25), KH2PO4 (1.25), and d-glucose (10).
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The preparation was transferred to the recording chamber and left there for at least 15 minutes before starting to record from the muscles.
The supernatant (soluble preparation) was transferred to a fresh tube, and the remaining pellet was resuspended in 7M urea, 2M thiourea, 1% CHAPS in 50 mM NH4HCO3, and 5 mM DTT at 60°C for 30 min (insoluble preparation).
DNA preparation was transferred into the tip and allowed to pass using gravity at room temperature.
After washing the preparation was transferred to the confocal microscope (LSM-5 Pascal Exciter, Zeiss, Germany).
A 0.1-ml aliquot of the OC preparation was transferred onto the slices.
An aliquot of the mitochondrial preparation was transferred to a disposable culture borosilicate glass tube (16 mm×100 mm).
Isometric force was measured after the preparation was transferred from relaxing to activating solution, by moving the stage of the inverted microscope.
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