Goat anti human serum albumin alb

For immunocytochemical staining, cells were initially fixed with 3.7% formaldehyde for 1 h and permeabilized with 0.25% Triton X-100 for 10 min at room temperature. After blocking with 1% bovine serum albumin in DPBS for 1 h, the cells were incubated overnight with primary antibodies, such as goat anti-human serum albumin (ALB; 1:200; Santa Cruz Biotechnology) and goat anti-hepatocyte nuclear factor 1-alpha (HNF-1α; 1:200; Santa Cruz Biotechnology) at 4 °C followed by incubation with CruzFluor™ 594 conjugated donkey anti-goat IgG (1:200; Santa Cruz Biotechnology) and CruzFluor™ 488 conjugated donkey anti-goat IgG (1:200; Santa Cruz Biotechnology) secondary antibody for 45 min at 37 °C, respectively. For counterstaining of cells nuclei 1 μg/ml 4′, 6-diamidino-2-phenylindole was used for 5 min and corresponding images were taken using a fluorescent microscope (Leica, Wetzlar, Germany). Untreated cells were also stained with corresponding antibodies to confirm that during immunostaining the detected signal in the differentiated cells was not due to background auto-fluorescence.

Immunocytochemical staining was performed by fixing cells with 3.7% formaldehyde for 30 min and permeabilized with 0.25% Triton X‐100 for 10 min at room temperature. After blocking with 1% bovine serum albumin (BSA) in DPBS for 1 h, cells were incubated with primary antibodies, such as goat anti‐Oct‐3/4 (1:200, Santa Cruz Biotechnology, CA), rabbit anti‐Sox‐2 (1:200, Santa Cruz Biotechnology), goat anti‐Nanog (1:200, Santa Cruz Biotechnology), goat anti‐human serum albumin (ALB, 1:200, Santa Cruz Biotechnology) and goat anti‐hepatocyte nuclear factor 1‐alpha (HNF‐1α, 1:200, Santa Cruz Biotechnology) for overnight at 4°C followed by incubation with CruzFluor™ 594 conjugated donkey anti‐goat IgG (1:200, Santa Cruz Biotechnology) or donkey anti‐rabbit IgG (1:200, Santa Cruz Biotechnology) secondary antibodies for 45 min at 37°C. The nuclei of cells were counterstained with 1 µg/ml 4′,6‐diamidino‐2‐phenylindole (DAPI) for 5 min and images were taken using fluorescence microscope (Leica, Wetzlar, Germany).