Journal: Journal of visualized experiments : JoVE

Article Title: Real-time In Vitro Monitoring of Odorant Receptor Activation by an Odorant in the Vapor Phase

doi: 10.3791/59446

Figure Lengend Snippet: Table of Materials

Article Snippet: Mix for transfection. table ft1 table-wrap mode="anchored" t5 caption a7 Name of Material/Equipment Company Catalog Number Comments/Description 0.05 % trypsin-EDTA Gibco 25300-054 0.05% Trypsin - EDTA (1x), phenol red - store at 4°C 100 mm cell culture dish BD Falcon 353003 100 mm x 20 mm cell culture dish 15 mL tube BD Falcon 352099 17 mm x 120 mm conical tubes 96-well plate Corning 3843 96 well, with LE lid white with clear bottom Poly-D-lysine coated Polystyrene Amphotericin Gibco 15290-018 Amphotericin B 250 µg/mL - store at 4°C centrifuge machine Jouan C312 Centrifuge machine with swinging bucket rotor for 15 mL Class II Type A/B3 fumehood NUAIRE NU-407-500 fumehood for cell culturing FBS Gibco 16000-044 Fetal Bovine Serum - store at −20°C GloSensor cAMP Reagent Promega E1290 GloSensor cAMP Reagent luminescent protein substrate - store at −20°C Incubator 37 °C; 5 % CO 2 Fisher Scientific 11-676-604 Incubator for cell culturing Lipofectamine 2000 reagent Invitrogen 11668-019 Lipofectamine 2000 Reagent 1mg/ml transfection reagent - store at 4°C Luminometer POLARstar OPTIMA BMG LABTECH discontinued 96 well plate reader for luminescence Mineral oil Sigma M8410 Solvent for odorants - store at room temperature Minimum Essential Medium (MEM) Corning cellgro 10-010-CV Minimum Essential Medium Eagle with Earle’s salts & L-glutamine - store at 4°C Penicillin/Streptomycin Sigma Aldrich P4333 Penicillin-Streptomycin solution stabilized with 10,000 U of penicillin and 10 mg streptomycin - store at −20°C pGlosensor Promega E2301 pGloSensor-22F cAMP luminescent protein plasmid - store at 4°C phase contrast microscope Leica 090-131.001 phase contrast microscope with x4, x10, x20 objectives RTP1S H. Matsunami lab - 100 ng/µL plasmid - store at 4°C Open in a separate window NOTE: We strongly advice to add an empty vector negative control (here Rho-pCI) and any positive control (OR known to respond to the tested odorant) to the experiment plan.

Techniques: Cell Culture, Transfection, Plasmid Preparation, Microscopy