N 2 hydroxylpiperazine n 2 ethanesulfonic acid

Pregnenolone, DHEA, adenosine 5′-triphosphate (ATP), dimethyl sulfoxide (DMSO), N-2-hydroxylpiperazine-N′-2-ethanesulfonic acid (HEPES), dithiothreitol (DTT), and Trizma base, were products of Sigma-Aldrich (St. Louis, MO, USA). PrimeSTAR® Max DNA polymerase was a product of Takara Bio (Mountain View, CA, USA). Oligonucleotide primers were synthesized by Eurofins Genomics (Louisville, KY, USA). Cellulose TLC plates and Ultrafree-MC 5000 NMWL filter units were from EMD Millipore (Billerica, MA, USA). Ecolume liquid scintillation cocktail was a product of MP Biomedicals, LLC. (Irvine, CA, USA). Carrier-free sodium [³⁵S]sulfate was from American Radiolabeled Chemicals (St. Louis, MO, USA). PAP[³⁵S] was synthesized using recombinant human bifunctional PAPS synthase based on a previously established procedure (Yanagisawa et al., 1998 (link)). All other chemicals were of the highest grade commercially available.

APAP, adenosine 5’-triphosphate (ATP), dimethyl sulfoxide (DMSO), dithiothreitol (DTT), and N-2-hydroxylpiperazine-N’-2-ethanesulfonic acid (HEPES) were products of Sigma Chemical Company. Morphine, tapentadol, and O-DMT were from Cayman Chemical. Cellulose thin-layer chromatography (TLC) plates were from Merck (EMD Millipore Corporation). Carrier-free sodium [³⁵S]sulfate was from American Radiolabeled Chemicals. 3’-Phosphoadenosine-5’-phospho[³⁵S]sulfate (PAP[³⁵S]) was synthesized using ATP and carrier-free [³⁵S]sulfate according to a previously established protocol [27 (link)]. X-Ray films were from Research Products International Corporation. Prime STAR^® GXL DNA Polymerase was a product of Clontech Laboratories, Inc. Protein molecular weight markers were from Bioland Scientific LLC. PCR kit was a product of G-Biosciences. QIAprep^® Spin Miniprep Kit was from QIAGEN. Ecolume scintillation cocktail was from MP Biomedical LLC. Glutathione Sepharose^TM was a product of GE Healthcare Bio-Sciences. All other chemicals were of the highest grades commercially available.