Mouse rat hgf quantikine elisa

For the detection of SASP factors in the serum (n = 4) or serum‐free conditioned culture medium of isolated HSC (n = 3) from young and old rats, the Proteome Profiler Rat Cytokine Array Kit (R&D Systems) was used according to the manufacturer's instructions. Analysis of serum samples revealed that 10 out of 29 factors showed a reaction on the membrane while only two factors could be detected on the membrane incubated with the culture medium. Visualization and documentation were performed with ChemiDoc Imaging System (BioRad) and protein dot intensities were analyzed with the AlphaView software (Proteinsimple). Furthermore, culture supernatants of HSC from young (n = 5) and old (n = 3) rats were analyzed with the Rat CXCL3/CINC‐2 alpha/beta Quantikine ELISA Kit (RCN200, R&D Systems) and the Rat IL6 Quantikine ELISA Kit (R6000B, R&D Systems) according to manufacturer's recommendations. Data from the cytokine array and ELISA analyses were normalized to cell number per area, which was assessed by counting the cells with the cellSens Dimension 1.16 software at three different positions of each culture dish (Olympus). To investigate the amount of HGF released by HSC after shear stress (n = 5) and stretching (n = 4) as well as by culture on different ECM proteins, culture supernatants were analyzed with the Mouse/Rat HGF Quantikine ELISA (R&D Systems).

Rat livers from young and old male Wistar rats were perfused via the portal vein using Krebs–Henseleit buffer essentially as described (Wettstein & Häussinger, 1997). The flow rate of the buffer was adjusted to the liver weight. Initially the liver was perfused with 2 ml/(min × g liver) and after 10 min the perfusion rate was increased to 6 ml/(min × g liver). The liquid pressure was continuously recorded during liver perfusion. The effluent was collected in fractions to enable quantification of HGF by the Mouse/Rat HGF Quantikine ELISA (R&D Systems). The liver perfusion experiments were approved by the Landesamt für Natur, Umwelt und Verbraucherschutz (81‐02.04.2018.A126).