Trophoblast cells were fixed in 4% paraformaldehyde solution for 15 min and blocked in 1% BSA in PBS for 30 min, followed by incubation overnight at 4°C with a primary antibody against Fyn (1 : 1000, Ab125016, Abcam, Cambridge, MA, USA). Afterwards, Cy3-conjugated secondary antibodies (1 : 1000, BA1032, Boster, Wuhan, China) were added for 1 hr at 37°C. The cell nuclei were stained with 4,6-diamino-2-phenyl indole (DAPI) (C1002, Beyotime Biotechnology, Shanghai, China) for 5 min, and the images were captured using a fluorescence microscope (BX53, Olympus, Tokyo, Japan).
4 6 diamino 2 phenyl indole dapi c1002
Manufactured by Beyotime
Sourced in China
4,6-diamino-2-phenyl indole (DAPI) (C1002) is a fluorescent dye used for nucleic acid staining in biological research. It binds strongly to adenine-thymine (A-T) rich regions in DNA, emitting blue fluorescence upon excitation.
Automatically generated - may contain errors
2 protocols using 4 6 diamino 2 phenyl indole dapi c1002
1
Fyn Protein Localization in Trophoblast Cells
2
Quantifying M1 and M2 Macrophages
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The sections were treated with rat anti‐F4/80 (Ab6640; 1:50; Abcam, Shanghai, China), mouse anti‐CD86 (NBP2‐44515, 1:50; Novus, Shanghai, China), and rabbit anti‐CD206 (Ab195191, 1:50; Abcam) primary antibodies (15 hours, 4℃) followed by staining with Cy3‐labeled goat anti‐rat IgG (Bs‐0293G‐CY3; 1:100, Bioss, Beijing, China), FITC‐labeled goat anti‐mouse IgG (BA1101, 1:100; Boster Biological Technology Co, Ltd, Wuhan, China), and FITC‐labeled goat anti‐rabbit IgG (BA1105, 1:10; Boster Biological Technology Co, Ltd) secondary antibodies (1 hour, 20℃), respectively. The sections were then treated with 4,6‐diamino‐2‐phenylindole (DAPI, C1002; Beyotime Biotechnology Co Ltd, Wuhan, China) to stain the nuclei (5 minutes, 20℃). The proportion of M1 and M2 macrophages was evaluated using the mean optical density (MOD) as determined by the ImageJ analysis software.
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