Ultrafree mc 5000 nmwl filter units

Pregnenolone, adenosine 5′-triphosphate (ATP), 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), Trizma base, and dithiothreitol (DTT) were from 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). Dpn I and the DNA ladder were products of New England Biolabs (Ipswich, MA, USA). QIAprep^® Spin Miniprep Kit was from QIAGEN (Germantown, MD, USA). Glutathione Sepharose was a product of GE Healthcare Bio-Sciences (Pittsburgh, PA, USA). Cellulose TLC plates and Ultrafree-MC 5000 NMWL filter units were from EMD Millipore (Billerica, MA, USA). X-ray films were sourced from Products International Corporation (Mt Prospect, IL, USA). Ecolume scintillation cocktail was purchased from MP Biomedicals, LLC. (Solon, OH, USA). Carrier-free sodium [³⁵S] sulfate was from American Radiolabeled Chemicals (St. Louis, MO, USA). [³⁵S]PAPS was synthesized from adenosine triphosphate (ATP) and carrier-free [³⁵S]sulfate using recombinant human bifunctional ATP sulfurylase/ adenosine 5’-phosphosulfate (APS) kinase. The [³⁵S]PAPS synthesized was adjusted to the required concentration and specific activity of 15 Ci/mmol at 1.4 mM by the addition of unlabeled PAPS^{17 (link)}. All other chemicals were of the highest grade commercially available.