Molecular Devices Corp

Fatty Acid Transporters. Figure 2 Quencher-based real-time fatty acid uptake assay with a fluorescently labeled FFA analogue (C1-Bodipy-C12). Predominantly protein-mediated fatty acid uptake by 3T3-L1 adipocytes (diamonds) was compared with diffusion-driven uptake by fibroblasts (squares) using the QBT Fatty Acid Uptake reagent (Molecular Devices Corp., CA, USA), which contains C1-Bodipy-C12 as substrate in conjunction with a cell impermeable quencher. Uptake kinetics was recorded using a Gemini fluorescence plate reader. Error bars indicate the standard deviations from 12 independent wells. RFU relative fluorescence units. 

Molecular Devices Corp PatchXpress 7000A 16-well 2000 Voltage control Low throughput... 

Molecular Devices Corp IonWorksHT 384-well 3000 Voltage control Low seal... 

Apparatus. Quantitation of color developed in the microwells was achieved by a Vmax Reader (Molecular Devices Corp., Palo Alto, California). Standard curve using semi-log curve fit and unknown calculations were accomplished by using the software program Softmax provided by the manufacturer. The Ultrawasher II (Dynatech Laboratories Alexandria, Virginia) was used to wash the wells. 

Microtiter plates (MaxiSorp-immunoplates, NUNC A/S, Roskilde, Denmark) were coated overnight at 23°C with the cohesin test samples (200 pl/well, 270 nM of miniCipC c, wild-type or mutated Coh2CBD t). The plates were blocked for 2.5 h with blocking solution (300 pl/well 3% (w/v) bovine serum albumin in TrisNC buffer) and washed three times with TrisNC buffer (300 pl/well). The cohesin-dockerin interaction was initiated upon addition of dockerin samples (200 pl/well, 94 nM of XynDocA c or XynDocS t), and the plates were incubated for 2.5 h. After five washes, the bound dockerins were detected by means of the fused-xylanase activity substrate solution (240 pl/well 2.9 mM / -nitrophenyl p-D-cellobioside) was added followed by incubation at 60 C. Optical density was detOTnined at 420 nm on a VERSAmax microplate reader (Molecular Devices Corp., Sunnyvale CA). 

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