Antibodies against tyrosinase

Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum (FBS) and antibiotics mixture were purchased from WELGENE (Gyeongsan-si, Gyeongsangbuk-do, Korea). Kojic acid (KA), phenylthiourea (PTU), mushroom tyrosinase, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), α-MSH and PD98059 were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA). Antibodies against tyrosinase, MITF, ERK, phospho-ERK (p-ERK) and anti-β-actin were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Peroxidase-labeled anti-rabbit and anti-mouse immunoglobulins were obtained from KOMA BIOTECH (Seoul, Korea). All other chemicals were purchased from Sigma-Aldrich. Anthocyanin powder of H. sabdariffa L. from Egypt was purchased from Shin Young Hub (Seoul, Korea).

Quercetin, tyrosinase (EC 1.14.18.1, 128 kDa), and l-DOPA were purchased from Sigma Aldrich (St Louis, MO, USA). Stock solutions of tyrosinase and l-DOPA (5.0 × 10⁻³ mol L⁻¹) were dissolved in 0.05 mol L⁻¹ sodium phosphate buffer, pH 6.8. Oleyl bromide, Na₂CO₃, and dimethyl sulfoxide (DMSO) were purchased from Sigma Aldrich. Arbutin was purchased from TCI (Tokyo, Japan), and PTU and Quercetin were purchased from Sigma Aldrich. PTU is a well-known tyrosinase inhibitor that is routinely used to inhibit pigment production in zebrafish. All other chemicals and reagents were of molecular biology grade and were commercially available. Antibodies against tyrosinase, TRP-1, TRP-2, MITF, PKA, p-PKA, CREB, and p-CREB were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). p-PKA and p-CREB antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). α-MSH, DMSO, sodium nitrite, and an antibody against β-actin were obtained from Sigma Chemicals (St. Louis, MO, USA). Horseradish peroxidase-conjugated goat anti-rabbit and anti-mouse antibodies were obtained from Invitrogen (Carlsbad, CA, USA).

Fraxinol (6-hydroxy-5,7-dimethoxychromen-2-one) was purchased from Cayman Chemical (Ann Arbor, MI, USA). Dulbecco’s modified Eagle medium (DMEM), fetal bovine serum, penicillin/streptomycin, and trypsin-ethylenediaminetetraacetic acid were purchased from HyClone (Logan, UT, USA), Corning, Inc. (Corning, NY, USA), Gibco (Grand Island, NY, USA), and Welgene (Gyeongsan-si, Republic of Korea), respectively. Dimethyl sulfoxide, α-MSH, NaOH, and MTT were obtained from Sigma–Aldrich (St. Louis, MO, USA). Antibodies against tyrosinase, TRP-1, TRP-2, MITF, and phosphorylated CREB were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). H89 cells were purchased from Abcam (Cambridge, UK). Enhanced chemiluminescence kits were obtained from GE Healthcare (Buckinghamshire, UK).

Gallacetophenone was purchased from Biosynth Carbosynth (Berkshire, UK), isolindleyin was purchased from ChemFaces (Wuhan, China), and ethyl caffeate was purchased from ALB Technology Limited (Kowloon, Hong Kong, China). MEthyl 3,4,5-trihydroxybenzoate, Ethyl 3,4,5-trihydroxybenzoate, KA, cycloheximide, MG-132, and chloroquine were purchased from Sigma-Aldrich (St. Louis, MO, USA). Antibodies against tyrosinase, TRP1, and TRP2 were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Antibody against MITF was purchased from Neomarkers (Carlsbad, CA, USA), and GAPDH was obtained from Cell Signaling Technology (Danvers, MA, USA).