Journal: Journal of visualized experiments : JoVE

Article Title: High Throughput Traction Force Microscopy Using PDMS Reveals Dose-Dependent Effects of Transforming Growth Factor-β on the Epithelial-to-Mesenchymal Transition

doi: 10.3791/59364

Figure Lengend Snippet: Materials

Article Snippet: NOTE: The mixture is mixed on a rotator at a speed slow enough for the mixture to flow back and forth during revolution to ensure complete mixing. table ft1 table-wrap mode="anchored" t5 Name Company Catalog Number Comments Plate GEL-8100 Nusil Technology GEL-8100 High Purity Dielectric, Soft Silicone Gel kit Dow Corning Sylgard 184 Silicone Encapsulant Clear 0.5 kg Kit Ellsworth Adhesives 184 SIL ELAST KIT 0.5KG curing agent Custom Cut Glass Hausser Scientific Company 109.6mm± × 72.8mm± × 1mm thickness Target 2TM Nylon Syringe Filter ThermoFisher Scientific F2513-4 96-well Stripwell Egg Crate Strip Holder Corning 2572 Polystyrene Universal Microplate Lid With Corner Notch Corning 3099 Ethyl alcohol Greenfield Global P016EA95 0.95 2-Propanol Sigma-Aldrich 190764 ACS reagent, ≥99.5% Surface Coating Sulfo-SANPAH Crosslinker Proteochem c1111-100mg Fibronectin bovine plasma Sigma-Aldrich F1141-1MG solution, sterile-filtered, BioReagent, suitable for cell culture PBS, 1X Wisent 319-005-CL pH 7.4, without calcium and magnesium DMSO Sigma-Aldrich 472301 Cell Culture DMEM, 1X Wisent 319-005-CL 4.5g/L glucose, with L-glutamine, sodium pyruvate and phenol red FBS (Fetal Bovine Serum) Wisent 080-150 Premium Quality, Endotoxin <1, Hemoglobin <25 HEPES Wisent 330-050-EL 1M, free acid Human Insulin Recombinant Wisent 511-016-CM USP grade Penicillin-Streptomycin Solution Wisent 450-201-EL 100 X, sterile filtered for cell culture L-Glutamine solution Wisent 609-065-EL 200mM solution, sterile filtered for cell culture Amphotericine B Wisent 450-105-QL 250μg/ml, sterile filtered for cell culture Recombinant Human TGF-β1 Peprotech 100-21 HEK293 Derived Acetic acid Sigma-Aldrich 537020 Glacial, ≥99.85% Cictric acid Sigma-Aldrich 251275 ACS reagent, ≥99.5% NMuMG ATCC CRL-1636 Mouse Mammary Gland Cell Line Sodium azide Fisher Schientific AC190385000 99%, extra pure, ACROS Organics Potassium hydroxide Sigma-Aldrich 221473 ACS reagent, ≥85%, pellets TritonX-100 Sigma-Aldrich X100 laboratory grade Bead Synthesis 1,1′-Dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate (Dil) Sigma-Aldrich 468495-100MG 97% Methyl methacrylate Sigma-Aldrich M55909-500ML contains ≤30 ppm MEHQ as inhibitor, 99% Inhibitor Remover Sigma-Aldrich 306312-1EA Prepacked column for removing hydroquinone and monomethyl ether hydroquinone Methacryloxylpropyl Terminated Polydimethylsiloxane Gelest DMS-R31 (25,000g/mol) Polydimethylsiloxane stabilizer, 25,000g/mol, 1,000 cSt 2,2′-Azobis(2-methylpropionitrile) (AIBN) Sigma-Aldrich 441090-25G 98% Hexane Sigma-Aldrich 296090-2L anhydrous, 95% Hexane, mixture of isomers Sigma-Aldrich 227064-1L anhydrous, ≥99% Whatman qualitative filter paper, Grade 1 Sigma-Aldrich WHA1001055 circles, diam.

Techniques: Stripping Membranes, Cell Culture, Recombinant, Derivative Assay