Evaluating 99mTc-RYM1 Binding in Lung Tissue

Ex vivo binding of ^99mTc-RYM1 to biological tissue was evaluated in lung tissue homogenate of CC10-IL-13Tg mice (n = 2) showing elevated basal MMP activity^{20 (link)}. Lung tissue (247 ± 85 µg, in duplicates was incubated with 3 nM of ^99mTc-RYM1 for 60 min at 37 °C with or without the preincubation of 20 µM of RYM. After tracer incubation, tissue homogenate was spin down and washed 3 times with PBS, before resuspension in protein lysis buffer, gamma-counting (Wizard2, PerkinElmer) and protein concentration was measured (Pierce BCA Protein Assay Kit, Thermo Fisher Scientific and Multiskan Ascent Microplate Photometer, Thermo Labsystems) in order to determine the amount of tissue used. All experimental protocols were approved by Yale University and VA Connecticut Institutional Animal Care and Use Committees.