Tgf β1 elisa kit

The bioactive substances present in the culture supernatant were analyzed using the TGF-β1 ELISA kit (#KE00002, Protein Tech, Rosemont, IL, USA), IL-8 ELISA kit (#KE00006, Protein Tech), and Sonic hedgehog (SHH) ELISA kit (#ab100639, Abcam) according to the manufacturer's protocol. The samples were analyzed on day 3 or day 4 when ballooning was first observed (Fig. S2). On day 11, PHH/HSC sheets were homogenized, and protein lysates were prepared. Lysates were analyzed using the p62 ELISA kit (#ADI-900-212, ENZO New York, NY, USA), and protein carbonyl ELISA kit (#STA-310, CELL BIOLABS, San Diego, CA, USA) according to the manufacturer's protocol. The BCA protein assay kit (#T9300A, Takara Bio) was used to determine the protein concentration in the lysates. The Human Albumin ELISA kit (#E88-129, Bethyl Laboratories, Inc., Waltham, MA, USA) and Urea assay kit (#DIUR-100, BioAssay Systems, Hayward, CA, USA) were used to assess hepatocyte function. The samples were analyzed on days 0, 1, 5, and 10.

P. gulae strains were used to infect Ca9-22 cells with or without administration of the IFN-α formulation, as described in the above section regarding RT-PCR. ELISAs were performed in accordance with previously described methods, with some modifications^{56 (link)}. Ca9-22 cells were stimulated with P. gulae strains in the presence or absence of IFN-α for 24 h. For the quantification of IL-1β, COX-2, IL-8, and TGF-β1 in cell lysate at each time point, sandwich ELISAs were performed using the Human IL-1β ELISA kit (Proteintech Group Inc., Rosemont, IL, USA), Human/mouse total COX-2 DuoSet IC ELISA (R&D Systems Inc., Minneapolis, MN, USA), IL-8 ELISA kit (Proteintech Group Inc.) and TGF-β1 ELISA kit (Proteintech Group Inc.), respectively, in accordance with the manufacturers’ instructions. Absorbance was measured at 450 nm, with correction to 550 nm, using a SH-1000 Lab microplate reader (Corona Electric, Ibaraki, Japan). All assays were performed in triplicate on three separate occasions (n = 9).