Thermo Fisher Scientific (Invitrogen)
Alexa Fluor 568-conjugated donkey anti-rabbit secondary antibody
A10042
N/A
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Cells cultured on sterile glass cover slips in 24-well plates were fixed with 4% paraformaldehyde (PFA) in PBS, washed and permeabilized with 0.1% Triton X-100 PBS. The cells were blocked with 1% goat serum for 1 hour and incubated with primary antibody overnight. After washing, the cells were incubated with secondary antibodies for 45 min. Then, the cover slips were mounted with Vectashield mounting medium containing DAPI (Vector Laboratories, Burlingame, USA). The immunostaining images were captured under a Leica fluorescent microscope (DM6000B, Leica, Germany). The results showed that Foxp1ECKO mutants exhibit greater AngII-induced cardiac fibrosis by increased numbers of PCNA/Vimentin (Vim) double positive cells by co-immunostaining in the heart compared with wild-type littermates.
Immunofluorescence
Mouse heart
1/200 dilution 4°C 30 minutes
1% goat serum
1/500 dilution room temperature 45 min incubation
Mounted with Vectashield mounting medium containing DAPI
Fluorescence microscope (Leica, Wetzlar, Germany)
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FIGURE 2D
The antibody works well as expected.
The antibody works.