Figure 5.

Eastern blot analysis of urine collected from EHBRs orally treated with GA using anti-3MGA MAb. On the photographs, base lines were added. On TLC plate, the following samples were spotted at the base line; lane 1, double spot of 6 µl of urine collected from EHBRs which were orally administered with GA as drinking water (1 mg/ml) and 2 µl of 3MGA (1 mM); lane 2, 2 µl of 3MGA (1 mM); lane 3, double spot of 6 µl of urine collected from EHBRs and 2 µl of compound 1 (1 mM); lane 4, 2 µl of compound 1 (1 mM); lane 5, 6 µl of urine collected from EHBRs. Then, the spots were spreaded out using H₂O/BuOH/AcOH (2:7:3), and the top line of the solvent was penciled. Photograph of TLC plate detected by UV absorption at 254 nm (left) was taken. On the lanes 2 and 4 used as positive control, the spots appeared were marked by pencil. The spots on the plate were transferred onto PES membrane, fixed, blocked, and strained by anti-3MGA MAb. Photograph of the membrane was taken (right). Rf values of 3MGA and compound 1 were 0.76 and 0.50 respectively.