Aim: To prepare and characterize monoclonal antibody (mAb) against human LSECtin (liver and lymph node sinusoidal endothelial cell C-type lectin) protein.
Methods: BALB/c mice were immunized with prokaryotically expressed human LSECtin protein. The splenocytes from the immunized mice were fused with murine myeloma cells (Sp2/0) and then the mAb-positive hybridoma cells were screened by indirect ELISA. Reaction of mAb to LSECtin antigen was characterized by Western blot, indirect immunofluorescent staining, immunohistochemical staining and FCM.
Results: Eight hybridoma cells secreting mAbs were established. The isotypes of the mAbs were IgG. Ascites titers were between 1:10(6) - 1:10(7). All the mAbs recognized human LSECtin protein on LSECtin-transfected 3T3 cells and six of the mAbs specifically recognized liver sinusoidal endothelial cells.
Conclusion: Eight anti-LSECtin mAbs have been obtained. The characterization of the mAbs indicate that they show fine specificity by Western blot, indirect immunofluorescent staining, immunohistochemical staining and FCM, which can provide a powerful reagent for the functional study of LSECtin.
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