N 3 dimethylaminopropyl n ethylcarbodiimide hydrochloride edc hcl

Methoxy-polyethylene glycol amine (mPEG-NH₂, Mw 5.000 Da) and mPEG propionic acid (Mw 5.000 Da) were purchased from JenKem Technology and Sigma-Aldrich, respectively. N-hydroxy succinimide (NHS) and N-(3-dimethylaminopropyl) -N-ethyl carbodiimide hydrochloride (EDC. HCl) were obtained from Sigma-Aldrich and used directly. All solvents used, 3-mercaptopropionic acid (3-MPA), dichloromethane (DCM), dimethylformamide (DMF), acetone, hydrogen peroxide (H₂O₂), hexane, and methanol were of analytical grade and used without further purification. Melphalan was purchased from Fisher Scientific (Milano, Italy). Ultrapure water, which was used for all the experiments, was provided by a Milli-Q water system (Millipore, Bedford, MA, USA). F12-K and trypsin (TrypLE Select Enzyme [1X]) were purchased from Gibco. Fetal bovine serum (FBS), penicillin-streptomycin, oxidative fluorescent dye, and dihydroethidium (DHE), were purchased from Sigma-Aldrich. DPBS and DMEM High Glucose were purchased from Lonza (Verviers, Belgium).

All reagents were used as received without further purification. MilliQ^TM water (Merck, Melbourne, Australia) was utilised, and organic solvents were of analytical grade. We acquired 1S,2S-Diaminocyclohexane (SSDACH, 98%), 4,7-dimethoxy-1,10-phenanthroline (47O₂Me₂Phen), 1,10-phenanthroline (Phen), 5-methyl-1,10-phenanthroline (5MePhen), 5,6-dimethyl-1,10-phenanthroline (56Me₂Phen), biotin, silver nitrate, N-(3-Dimethylaminopropyl)-N’-ethylcarbodiimide hydrochloride (EDC-HCl), and N-hydroxysuccinimide (NHS) from Sigma-Aldrich, Sydney, NSW. Potassium tetrachloroplatinate (K₂PtCl₄) was obtained from Precious Metals Online. Potassium iodide (KI) was purchased from Merck, Melbourne, Australia. Hydrogen peroxide (H₂O₂, 30%) was obtained from VWR chemicals, Philadelphia, USA. Acetonitrile (ACN), dimethyl formamide (DMF), diethyl ether, methanol (MeOH), and dimethyl sulfoxide (DMSO) were purchased from Chem-Supply, Adelaide, Australia. Sep-Pak^® C18 columns were acquired from Waters, Australia. For ICP-MS experiments, the certified reference standard used was purchased from High-Purity standards, North Charleston, United States. Ultra-pure HNO₃ (69%) was obtained from Choice Analytical, Sydney, Australia. Deuterated solvents used for NMR experiments (d₆-dimethyl sulfoxide (DMSO-d₆, 99.9%) and deuterium oxide (D₂O, 99.9%) were acquired from Novachem, Melbourne, Australia.

Dulbecco´s Modified Eagle´s Medium with Ham’s F-12 Nutrient Mixture (DMEM-F12) with L-glutamine was purchased from CORNING (New York, NY, USA). Cervical cancer HeLa cells were obtained from ATCC (Manassas, VA, USA). 3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was supplied by Sigma-Aldrich (St. Louis, MO, USA). PTX was purchased by MedChemExpress (Monmouth Junction, NJ, USA). For G4-PTX-R8 synthesis, the dendrimer of fourth generation containing ethylenediamine core groups and surfaced amino groups (G4 PAMAM) was acquired from Dendritech (Midland, MI, USA). N-hydroxysuccinimide and N-(3-Dimethylaminopropyl)-N-ethyl carbodiimide hydrochloride (EDC. HCl, 98%) were supplied by Sigma Aldrich Chemicals (St. Louis, MO, USA). Finally, the cellulose dialysis membrane (MWCO 3.5 KDa) was obtained from Spectrum Laboratories, Inc. (Dominguez, CA, USA). All other commercially purchased solvents and chemicals were of analytical grade.
The reagents required for CRISPR-Cas9 technology (single guide RNAs (crRNAs), ATTO-labelled tracrRNA, and Cas9) were obtained from IDT (IA, Coralville, IA, USA); Buffer R and the primers for Bmal1 were purchased from Invitrogen (Carlsbad, CA, USA). The ApoAlertTM Caspase-3 Colorimetric assay kit was purchased from Clontech Lab (Orange, CA, USA), and Caspase-Glow^® 9 Assay from Promega (Madison, WI, USA).