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The blocking buffer was decanted from each container.
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ChonBlock™ blocking buffer was procured from Chondrex, Inc. (Redmond, WA).
After the coating buffer was decanted, 200 μL of blocking buffer (phosphated-buffered saline [PBS], 0.05% Tween 20, 5% dry milk) was added to each well, and the wells were incubated at room temperature for at least 30 minutes.
The decellularization buffer was decanted daily and replaced with fresh decellularization buffer.
After immobilization, the buffer was decanted and the beads were used for transesterification without further modification.
The assay buffer was decanted and the tissues were bleached in 95% EtOH and photographed.
The buffer was decanted and 6 μg of total protein in preactivation buffer was applied onto each spot.
After a short centrifugation, washing buffer was decanted and 70%% ethanol added for a 5 minute incubation at room temperature.
The wash buffer was decanted and the standards, assay buffer, or serum samples were mixed with serum matrix in each well.
Osmium fixative was removed and sperm washed thrice with 0.1 M cacodylate buffer by keeping on ice for 15 min, followed by short spin and buffer was decanted.
The buffer was decanted, and the cells were resuspended in OS solution 2 [20 mM Tris-HCl (pH 8.0), 2.5 mM EDTA] using the same volume as before.
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