Western blotting method for the immunostaining detection of glucuronides of glycyrrhetic acid using anti-glycyrrhizin monoclonal antibody

A method for detecting glucuronides of glycyrrhetic acid using Western blotting was investigated. Glucuronides of glycyrrhetic acid separated by silica gel TLC were transferred to a polyvinylidene difluoride membrane. The membrane was treated with sodium periodate solution followed by bovine serum albumin, resulting in a glucuronides of glycyrrhetic acid-BSA conjugate. Individual spots were stained by monoclonal antibody against glycyrrhizin. Immunostaining of glucuronides of glycyrrhetic acid was more sensitive compared to other staining methods. The newly established immunostaining method can be expanded to the distribution of glucuronides of glycyrrhetic acid in the plant body.