Figure 2: Using immunoprecipitation to confirm the presence of TLR4 in HGF. HGF cell lysate was added to anti-Toll-like receptor 4 (TLR4) rabbit polyclonal antibody and incubated with gentle rocking overnight at 4°C. Protein L-Sepharose was also added. The sample was loaded on SDS-PAGE gel (12–15%). The sample was then analyzed by silver staining. The TLR4 protein was visualized as a 96-kDa band.TLR4 in HGF was detected by immunoprecipitation with an antibody against TLR4. Lane 1, molecular-weight (MW) standard; Lane 2, Protein L-Sep only; and Lane 3, Protein L-Sep +HGF+ TLR 4 antibody. 1-DE patterns were visualized by silver-staining.