Fitc labeled goat anti mouse igg a0568

To identify isolated primary calf hepatocytes, we applied immunofluorescence to detect the expression of cytokeratin-18 (CK-18) a commonly used marker protein of hepatocytes (Jiang et al., 2013) (link). After 72 h, the cells were washed with PBS and fixed with 4% paraformaldehyde for 30 min at room temperature. To block nonspecific binding, hepatocytes were incubated with 3% Albumin Fraktion V (Carl Roth, Karlsruhe, Germany), 5% normal goat serum (Sigma, Darmstadt, Germany), and 0.5% Triton in PBS (PAA, Cölbe, Germany) for 30 min at room temperature. The cells were then exposed to a mouse monoclonal CK-18 antibody (ab668, 1:400; Abcam, Cambridge, UK) at 4°C in a humidified chamber overnight. The cells were then rinsed 4 times with PBS and incubated with fluorescein isothiocyanate (FITC)-labeled goat anti-mouse IgG (A0568, 1:500; Beyotime, Shanghai China) for 1 h at room temperature. After the cells were washed 3 times and covered with a glass coverslip, images were acquired with a laser confocal microscope (LSM 5 Pascal, Zeiss, Oberkochen, Germany). The results showed that the cytoplasm of more than 95% of the cells contained CK-18 (Supplemental Figure S1; https: / / doi .org/ 10 .3168/ jds .2019 -17920).