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After cooling the product to room temperature, the remaining thionyl chloride was removed by repeated additions of toluene under vacuum conditions

The lower pH helps protonate the peptide, ramping up its net positive charge and making it much more friendly with the water molecules

Hum Reprod Update 10:107117 Jones T, Ho JR, Gualtieri M, Bruno GJ, Chung K, Paulson RJ (2018) Clomiphene stair-step protocol for PCOS

Clinical trials extending 68 weeks show acceptable safety profiles with no unexpected serious adverse events

At 48 h post-transfection, cells were harvested in ice-cold PBS using Trypsin-EDTA (Lonza, Switzerland), centrifuged at 1000 g for 3 min, and washed with ice-cold PBS