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Plates were shaken for 5 min and then the blue solution was transferred to a 96-well plate; optical densities were read at 570 nm in a multiwell spectrophotometer (TECAN Infinite® 200).
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A solution of methylene blue (0.01% w/v) was added to the cell suspension and after one minute this solution was transferred to a Neubauer-improved chamber in order to count the cells.
A fiber in pCa8.0 relaxing solution was transferred to pCa8.0 rigor solution (0 mM ATP) including 1 unit/mL apyrase [21], and then transferred to pCa4.5 rigor solution.
The upper solution was transferred into an HPLC vial.
The resulting clear solution was transferred to a new tube.
Each hexane solution was transferred to a chromatography vial.
Organs were incubated with X-gal staining solution (5 mM potassium hexacyanoferrate, 5 mM potassium hexacynoferrate trihydrate, 1 mg/ml X-gal in rinse buffer) for 1 hour at 37°C or room temperature with gentle agitation until the appearance of blue coloration and then organs in X-gal staining solution were transferred to 4°C refrigerator overnight.
The reconstituted solutions were transferred to NMR tubes.
The solutions were transferred to an ultrasonic bath.
It contained protein since the color of Coomassie Blue solution was turned into blue.
Methylene blue solution was used to aid visualization by blue color.
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