Tris buffer

G6P was purchased from USB Corporation, unless otherwise specified. The F₄₂₀ cofactor with a tail length of 6-9 γ-glutamate residues was prepared as previously described [8 (link)]. Activated carbon, ATP disodium salt (98%), sodium citrate, magnesium chloride and Tris buffer were purchased from Fisher Scientific. D₂O, deuterium chloride, D-Glucose and D-Glucose-1-d₁ (98 atom % D) were purchased from Sigma Aldrich. Hexokinase was purchased from Worthington Biochemical Corporation. The glucose-6-phosphate assay kit was purchased from ScienCell Research Laboratories.

PMMA sheets (T_g = 105°C), 1.5 mm and 0.175 mm thick, were purchased from Good Fellow (Berwyn, PA). COC 6017 (T_g ≈ 178°C), 5010 (T_g ≈ 108°C) and 8007 (T_g ≈ 78°C; 0.13 mm) sheets were purchased from TOPAS Advanced Polymers (Florence KY). Si <100> wafers were secured from University Wafers (Boston, MA). Tripropylene glycol diacrylate (TPGA), trimethylolpropane triacrylate (TMPA), Irgacure 651 (photo-initiator), 50% potassium hydroxide (KOH), hydrochloric acid (HCl) and potassium chloride (KCl) were obtained from Sigma-Aldrich (St. Louis, MO). The anti-adhesion monolayer of (tridecafluoro – 1,1,2,2 – tetrahydrooctyl) tricholorosilane (T-silane) was purchased from Gelest, Inc. Tris buffer (pH = 8.0) was obtained from Fisher Scientific (Houston, TX). All dilutions were performed using 18 MΩ/cm milliQ water (Millipore).

Ethylacetate (EtOAc), methanol (MeOH), hydrochloric acid (HCL), ethylenediaminetetracetic acid (EDTA), sodium chloride (NaCL), tris buffer, and sodium hydroxide (NaOH) were purchased from Fisher Scientific (Hampton, NH). Ammonium bicarbonate, (+)-catechin, and trichloroacetic acid (TCA), arecoline hydrobromide, and guvacine hydrochloride were purchased from Sigma Aldrich (St. Louis, MO). Sodium deoxycholate (SDC) was purchased from Thermo Scientific (San Jose, CA). Guvacoline hydrobromide, N-nitrosoguvacoline, arecaidine hydrobromide, and arecodine-d₅, hydrobromide salt were purchased from Medical Isotopes (Pelham, NH). AN, betel leaves and slaked lime were purchased from a local grocery store in Honolulu.

ALL PT-3 cells (2×10⁶) were cultured in MEMα with either VAY736 (20 μg/mL), recombinant BAFF (200 ng/mL), or combination for 72 hours. Indicated cells were washed with PBS, followed by lysis with 9 M urea buffer (Fisher Chemicals) in Tris Buffer (Fisher Bioreagents). Supernatant was collected after centrifuging lysed cells at 14000 × g for 10 minutes at 4⁰C, and protein concentration was estimated using Thermo Scientific Nanodrop Lite Spectrophotometer. Protein samples (30 μg) were run on an SDS-polyacrylamide gel electrophoresis and transferred to a nitrocellulose membrane (BioRad, USA). The membrane was blocked for 1 hour with 5 % nonfat dry milk prepared in Tris Buffer Saline (TBS) containing 0.1% Tween 20 (Fisher Scientific). Membranes were probed with the primary antibodies overnight at 4⁰C, and with secondary antibodies for 1 hour at room temperature. Respective mouse or rabbit HRP-conjugated secondary antibodies were used to detect specific protein bands. Blots were developed using Clarity Western ECL substrate (BioRad, USA).