T butoxycarbonyl gln ala arg 7 amino 4 methylcoumarin boc qar amc

HEK293 cells were purchased from American Type Culture Collection (Manassas, VA). These cells were cultured in high glucose Dulbecco’s Modified Eagle’s Medium (DMEM) with 10% foetal bovine serum, 2 mM L-glutamine, 100 IU/ml penicillin and 100 µg/ml streptomycin. Serum-free media HCELL-100 was purchased from WISENT (St-Bruno, Canada). Poly-L-lysine coated coverslips were purchased from Corning (Bedford, MA). Anti-V5, Anti-V5 HRP and Anti-V5 FITC-linked monoclonal antibodies were purchased from Invitrogen (Waltham, MA). HRP-linked Anti-GAPDH rabbit monoclonal antibody was purchased from Cell Signaling Technology (Danvers, MA). Goat polyclonal anti-Hemojuvelin antibody and t-butoxycarbonyl-Gln-Ala-Arg-7-amino-4-methylcoumarin (Boc-QAR-AMC) were purchased from R&D Systems (Minneapolis, MN). Lipofectamine 3000 was purchased from Invitrogen (Carlsbad, CA). Centrifugal filters were purchased from Merck Millipore (Cork, Ireland). Lysis buffer (1% Triton, 50 mM Tris, 150 mM NaCl, 5 mM EDTA) was supplemented with protease inhibitor from Roche (Mannheim, Germany). Protein A/G PLUS-agarose beads were purchased from Santa-Cruz Biotechnology (Dallas, TX).

HEK293 and HepB cells were purchased from American Type Culture Collection (Manassas, VA). Hek293 cells were maintained in high glucose Dulbecco’s Modified Eagle’s Medium (DMEM) and Hep3B were cultured in Eagle’s Minimum Essential Medium (EMEM), both medium were supplemented with 2 mM L-glutamine, 100 IU/ml penicillin, 100 μg/ml streptomycin and 10% foetal bovine serum. HCELL-100 was acquired from WISENT (St-Bruno, Canada). Anti-V5 antibody was purchased from Invitrogen (Waltham, MA). Anti-HA antibody was purchased from Roche (Mannheim, Germany). HRP-linked Anti-GAPDH rabbit monoclonal antibody was purchased from Cell Signaling Technology (Danvers, MA). t-butoxycarbonyl-Gln-Ala-Arg-7-amino-4-methylcoumarin (Boc-QAR-AMC) was purchased from R&D Systems (Minneapolis, MN). Lipofectamine 3000 was purchased from Invitrogen (Carlsbad, CA). Centrifugal filters were purchased from Merck Millipore (Cork, Ireland). Lysis buffer (1% Triton, 50 mM Tris, 150 mM NaCl, 5 mM EDTA) was supplemented with protease inhibitors from Roche (Mannheim, Germany). Protein A/G PLUS-agarose beads were purchased from Santa-Cruz Biotechnology (Dallas, TX). Anti-V5-tag mAb magnetic beads were purchased from MBL (Woburn, MA). Pierce MS-Grade trypsin was purchased from Thermo Fisher Scientific (Waltham, MA).