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Definity Perflutren lipid microspheres (Lantheus Medical Imaging, North Billerica, MA, USA) were shaken using a Lantheus device for 45 s at 3000 r.p.m.
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The mixtures were shaken in a c24k refrigerated incubator shaker (NJ, USA) at room temperature for 60 min.
The vials were shaken at room temperature to extract steroids into aqueous phase and steroid levels were estimated using a liquid scintillation counter (Beckman, USA).
The solutions were shaken for 30 min at 37°C at 450 cycles per min in the vertical position using an Eppendorf mixer 5432 (Eppendorf, USA).
All the treatments were shaken at Speed 3 on the mixer (Cole-Parmer, USA) at 25°C.
After incubation, the plates were shaken thoroughly for 1 minute, and the absorbance was measured at 450 nm and 630 nm using an ELISA plate reader (BioTek, Winooski, VT, USA).
The plates were shaken for 5 min with a plate shaker and the absorbance was measured at 570 nm using a Bio-Rad ELISA plate reader (Bio-Rad, CA, USA).
The flasks were shaken at 26˚C for 2 hours [ 58], and the electrical conductivity of the solution (EC1) was measured using an IQ170 electrical conductivity meter (IQ Scientific Instruments, San Diego, CA, USA).
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