Use of asiatic acid in preparing medicine
A kind of asiatic acid, a technology for preparing medicine, applied in the field of medicine preparation of asiatic acid, can solve problems such as unreported
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[0048] Asiatic acid acts on rat hepatic stellate cells, prepares protein samples, Western blot:
[0049] 1. HSC-T6 cells 1×10 4 Spread 6-well plates, 5% CO 2 , 37°C, after 24 hours, add asiatic acid (purchased from SIGMA, USA) at concentrations of 5 μM, 10 μM, 20 μM, and 30 μM, add TGFβ 1 ng / ml after 2 hours, continue to cultivate for 24 hours, and extract cell protein by conventional methods. For protein content determination, store at -70°C for later use or do Western blot directly.
[0050] 2. The Western blot method is completely based on the second edition of "Molecular Cloning Experiment Guide" translated by Jin Dongyan, Li Mengfeng, etc. See pages 880-885 for protein acrylamide gel electrophoresis, and see pages 888-893 for protein transfer from SDS acrylamide gels to PVDF membranes.
[0051] 3. Wash the membrane with TBST after transfer. Block with 5% skimmed milk powder-TBST blocking solution at room temperature for 60 min; wash the membrane with TBST. Add the pr...
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