To generate metastases, 5-6 week old female NIH-III mice were anesthetized by intraperitoneal (i.p.) injection of ketamine (100 mg/kg) and xylazine (6 mg/kg), and then 2×105 MDA-MB-231-Luc2 cells suspended in 100 µl of PBS were injected into the left ventricle of each mouse. Successful intra-cardiac injections were confirmed by immediate whole body bioluminescence imaging. BAPN intraperitoneal injections at a dose of 100 mg/kg in 100 µl PBS, were initiated either 1 day prior, on the same day as, or 7 days after intracardiac tumor cell injection.

Enzyme activlty was measured by a peroxidase-coupled fluorescence assay. Reaction mixtures contained 2.5 mM diaminopentane, 10 ug o horseradish peroxldase, 0.7 mM homovanllllc acid and lysyl oxidase in 50 mM sodium borate buffer. Fluorescence was continuously monitored at excitation wavelength of 315 nm and emission wavelength of 425 nm. Lysyl oxidase was incubated with various amounts (10^-8 - 10^-4 M) of BAPN at 37°C C for 2 h in 16 mH potassium phosphate.