Abstract: Objective: To establish hybridomas that produces anti-death receptor 5 mAbs and checks surface expression of death receptor on cell lines. Methods: BALB/c mice were immunized with sDR5 in CFA. The spleen cells were fused with the myeloma SP2/0-Ag 14 in the presence of 50% polyethylene glycol. Surface expression of TRAIL receptor was determined by flow cytometric analysisby measuring the binding of anti-DR5 mAb. Results: The expression level of the different cell surfaceDR5 were 95% on SMMC7721 cells, 40% on HepG2 cells and 20.3% on HL7702 cells. Conclusions:These results demonstrate that anti-DR5 mAb is able to specifically bind to DR5 and that DR5 is ex-pressed at high level on SMMC7721 cell lines and middle level on HepG2 cell lines. The expression of DR5 on HL-7702cell lines is low. The DR5 specific mAb described here will serve as a valuable tool for studying the biology of TRAIL the receptors. The anti-DR5 mAb generated in the present study maybe useful for investigating DR5 expression in tissues or cell lines.