N benzoyl l tyrosine ethyl ester

Sodium phosphate, sodium chloride, triazine dye Cibacron Blue 3GA and the protease substrates N-benzoyl-L-tyrosine ethyl ester (BTEE) and N^α-benzoyl-L-arginine ethyl ester (BAEE) were obtained from Sigma-Aldrich (Darmstadt, Germany). Potassium thiocyanate (KSCN) was obtained from Carlo Erba (Chaussée du Vexin, France). Hydrogen chloride (HCl) was obtained from Scharlab (Barcelona, Spain). All other chemicals were of analytical grade and purchased from Merck (Darmstadt, Germany). The structures were created using ChemDraw Ultra 12.0 (CambridgeSoft, Cambridge, MA, USA, www.cambridgesoft.com).

l-Cysteine ethyl ester hydrochloride was purchased from Tokyo Kasei Co. (Tokyo, Japan). α-Chymotrypsin (EC 3.4.21.1) type II from bovine pancreas, 3×crystallized from 4× crystallized chymotrypsinogen, dialyzed essentially salt-free and prepared as lyophilized powder, which was 54 U/mg protein determined at 25 °C and pH 7.8 with N-benzoyl-l-tyrosine ethyl ester as a substrate, lysozyme from chicken egg white, l-arginine ethyl ester dihydrochloride and ProteoMass™ peptide MALDI-MS calibration kit were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA). 2,5-Dihydroxybenzoic acid, DMSO, trifluoroacetic acid (TFA) and urea were purchased from Kanto Chemical Co. (Tokyo, Japan). Iodoacetamide (IAM) and 5,5-dithiobis (2-nitrobenzoic acid) (DTNB) were obtained from Nacalai Tesque Inc. (Kyoto, Japan). 2,4,6-Trinitrobenzenesulfonic acid sodium salt dihydrate was obtained from Wako Pure Chemical Industries Ltd. (Osaka, Japan). Standards for size exclusion chromatography were from Bio-Rad (Hercules, CA, USA). All other regents used were of analytical grade.

N-benzoyl-L-tyrosine ethyl ester (BTEE), Na-Benzoyl-Larginine 4-nitroanilide hydrochloride (BAPNA), sucralose (≥98.0% HPLC), and Azoxymethane (AOM) were purchased from Sigma-Aldrich (St. Louis, MO, USA). 4-Nitrophenyl b-D-glucopyranoside was purchased from BBI Life Sciences. DSS (MW: 36–50 kDa) was obtained from MP Biomedical (Solon, OH, USA). The antibodies used in this study were anti-TLR4 (19811-1-AP, Proteintech), anti-VEGF (19003-1-AP, Proteintech), anti-STAT3 (10253-2-AP, Proteintech), anti-Phospho-Stat3 (Tyr705) (#9145, Cell Signaling Technology), anti-MyD88 (#4283, Cell Signaling Technology), anti-TRAF6 (YT4720, Immunoway), anti-inhibitor of NF-κB alpha (IκBα) (#4814, Cell Signaling Technology), and anti-Occludin (13409-1-AP, Proteintech), as well as anti-GAPDH, anti-β-Actin, goat anti-rabbit IgG, and goat anti-mouse IgG from ZSGB-BIO Co. Ltd. (Beijing, China). All other reagents used were of analytical grade.