Elastase from porcine pancreas type 4

The extraction and isolation solvents were purchased from Junsei Chemical Co., Ltd., Tokyo, Japan and Fisher Scientific company, Hampton, NH, USA. Chemicals for antioxidant assays were acquired from Fujifilm Wako Pure Chemical Corporation, Osaka, Japan. Elastase from porcine pancreas Type IV, tyrosinase from mushroom lyophilized powder, N-Succinyl-Ala-Ala-Ala-p-nitroanilide (SANA), L-tyrosine, oleanolic acid, vanillin, myricetin, and all buffer components were acquired from Sigma-Aldrich, St. Louis, MO, USA.
Pure momilactones A, B, and tricin were isolated previously from rice husk by our laboratory [12 (link)]. Briefly, the ethyl acetate (EtOAc) extract of 7 kg Koshihikari husks was separated by open column chromatography over silica gel with the mobile phase as combinations of hexane and EtOAc. Momilactones A and B were isolated from the eluant hexane:EtOAc (8:2) by a repeated column chromatography while tricin (a yellow powder) was purified from the last fractions of the eluant hexane: EtOAc (7:3). The identification of such pure compounds is described in detail in the following parts.

Methacholine (MCh) was obtained from ICN Biomedicals (Zoetermeer, the Netherlands). Alexa Fluor® 488 Phalloidin was purchased from Life technologies. Mouse anti-E-cadherin was obtained from BD Biosciences (Bedford, MA, USA), and Cy3-conjugated secondary antibody was purchased from Jackson ImmunoResearch (West Grove PA, USA). Elastase from porcine pancreas Type IV and chloroquine were received from Sigma-Aldrich (Zwijndrecht, The Netherlands).