赛默飞世尔Thermo Fisher INDICATOR F

Use of Spectral Fluorescence Resonance Energy Transfer to Detect Nitric Oxide‐Based Signaling Events in Isolated Perfused Lung

) Petri dish with coverslip bottom (e.g., Delta T dish, Bioptics) Perfusate and airway lines: polyethylene PE‐50 tubing

Fluorescence Spectroscopy

., Alexa Fluor 488 carboxylic acid, succinimidyl ester [Invitrogen]; FITC [Thermo Fisher Scientific]; rhodamine‐maleimide [Biotium]) DMSO, anhydrous grade

Stripping Western Blots

μM calyculin A, 2 mM PMSF, and 10 μg/ml aprotinin). 4) Lyse for 20 min on ice and clarify lysates by centrifugation at 16,000 x g for 10 min at 4 oC 5) Preclear lysates with 30 μl of a 1:1 slurry of protein G-sepharose (Fast-Flow; Pharmacia