Tris hydroxymethyl aminomethane trizma base

Medium 199 containing Earle’s salt and L-glutamine (M199), Dulbecco’s Modified Eagle Medium (DMEM-F12 medium), Tris-Ethylenediaminetetraacetic (EDTA), Bovine serum albumin (BSA), collagenase Type IA, from Clostridium histolyticum, Trypan Blue, tris(hydroxymethyl)aminomethane (Trizma base), and 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) were obtained from Sigma Chemical Company (St. Louis, MO, USA). TMRE (tetramethylrhodamine ethyl) was purchased from Fluka Company. hCG-Pregnyl 3,000 IU/mg (human chorionic gonadotropin) was from Organon (Roseland, New Jersey, USA). Power SYBR Green PCR Master Mix was purchased from Applied Biosystems (Thermo Fisher Scientific, Waltham, MA, USA), while qPCR primers were from Integrated DNA Technologies (Coralville, Iowa, United States). Anti-testosterone-11-BSA serum No. 250 was kindly supplied by Gordon D. Niswender (Colorado State University, Fort Collins, CO). Testosterone was from New England Nuclear (Brisel, Belgium). Active charcoal—Norit A—was obtained from Serva (Heidelberg, Germany).

Regarding chemicals, 0.01 M phosphate buffered saline (PBS) powder sachets (0.01 M NaH₂PO₄, 0.138 M NaCl, 0.0027 M KCl), Tris(hydroxymethyl)-aminomethane (Trizma base, ≥99.9%), sodium chloride (NaCl ≥ 99.5%), sodium hydroxide pellets (NaOH, ≥97%), mercuric chloride (HgCl₂, ≥99.5%), L-cysteine (Cys, ≥98% purity), and L-glutathione reduced (GSH, ≥98% purity) were purchased from Sigma-Aldrich (St. Louis, MO, USA). HCl (plasma pure 34–37%) was purchased from SCP Science (Baie D’Urfe, QC, Canada), while CH₃HgOH (aqueous 1.0 M solution) was obtained from Alfa Aesar (Haverhill, MA, USA). Arsenobetaine bromide (AB) was synthesized according to a previously published procedure, and its recrystallization from ethanol yielded white crystals with a melting point of 225 °C (literature: 227 °C) [53 (link)].