Immunofluorescence was used to detect F4/80, CD45.1, and CD45.2 as described by us previously [13 (link)]. Briefly, frozen livers were cut into 8 μm sections, fixed in 4% formalin for 10 minutes, followed by blocking in 10% goat serum. The sections were then incubated with Alexa Fluor-488 anti-mouse CD45.1 antibody (Biolegend) diluted 1:100, Alexa Fluor-488 anti-mouse CD45.2 antibody (Biolegend) diluted 1:100, or rat anti-F4/80 antibody (Bio-Rad, Hercules, CA) diluted 1:500. The sections were then incubated with goat anti-rat secondary antibody conjugated to Alexa Fluor-594 diluted 1:500 (Thermo-Fisher Scientific).
Goat anti rat secondary antibody conjugated to alexa fluor 594
Manufactured by Thermo Fisher Scientific
The Goat anti-rat secondary antibody conjugated to Alexa Fluor-594 is a laboratory reagent designed for use in various immunoassay techniques. It consists of a goat-derived antibody that specifically binds to rat primary antibodies, coupled with the Alexa Fluor-594 fluorescent dye. This product can be used to detect and visualize target proteins or antigens in samples when used in conjunction with a rat primary antibody.
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3 protocols using goat anti rat secondary antibody conjugated to alexa fluor 594
1
Immunofluorescence Analysis of Liver Cells
2
Immunofluorescence Analysis of Liver Cells
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Immunofluorescence was used to detect F4/80, CD45.1, and CD45.2 as described by us previously [13 (link)]. Briefly, frozen livers were cut into 8 μm sections, fixed in 4% formalin for 10 minutes, followed by blocking in 10% goat serum. The sections were then incubated with Alexa Fluor-488 anti-mouse CD45.1 antibody (Biolegend) diluted 1:100, Alexa Fluor-488 anti-mouse CD45.2 antibody (Biolegend) diluted 1:100, or rat anti-F4/80 antibody (Bio-Rad, Hercules, CA) diluted 1:500. The sections were then incubated with goat anti-rat secondary antibody conjugated to Alexa Fluor-594 diluted 1:500 (Thermo-Fisher Scientific).
3
Immunofluorescence Liver Staining Protocol
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Immunofluorescence was used to detect F4/80 and CD68 as described previously 20 . Briefly, 8 µm sections were cut from frozen livers and fixed for 10 minutes in 4% formalin. The sections were then incubated in blocking buffer (10% goat serum; 1 hour) followed by incubation with either rat anti-F4/80 antibody (Bio-Rad) diluted 1:500 or rat anti-CD68 antibody (Bio-Rad) diluted 1:500 overnight at 4°C. After washing, the sections were incubated with goat anti-rat secondary antibody conjugated to Alexa Fluor 594 for 1 hour (diluted 1:500, Thermo Fisher Scientific). Proliferating cell nuclear antigen (PCNA) was detected as described previously 20 .
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