Western blot analysis of CCL2 / MCP-1 on different lysates with Rabbit anti-CCL2 / MCP-1 antibody (HA500267) at 1/5,000 dilution. Lane 1: RAW264.7 cell lysate Lane 2: RAW264.7 treated with 100nM TPA overnight then 100ng/mL LPS for 7 hours add 1μg/mL BFA for last 3 hours cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 16 kDa Observed band size: 16 kDa Exposure time: 10 seconds; ECL: K1801; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA500267) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.