2.12. Western Blot analysis of iNOS and COX-2

Cell lysates were prepared in lysis buffer, and the protein concentrations were determined by Bio-Rad protein assay kit (Richmond, CA, U.S.A.). Samples of protein (50 μg) were electrophoresed using 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and then transferred to nitrocellulose membrane. The iNOS, COX-2 were assayed by specific antibodies (Santa Cruz Biotech, Santa Cruz, CA, U.S.A. Cell Signaling Technology). An enhanced chemiluminescence detection kit was used in immunodetection (Amersham, Piscataway, NJ).