MH-S cells were seeded into 15-mm glass bottom dishes (Nest Biotechnology, Jiangsu, China) at a density of 2 × 105 cells. After fixation with 4% paraformaldehyde, cells were permeabilized with 0.5% Triton X-100, blocked in blocking buffer, and incubated overnight with an anti-α1-AR antibody (1:100; Abcam, Cambridge, UK) at 4°C. The following day, cells were incubated with a fluorochrome-conjugated secondary antibody [1:500; Cell Signaling Technology (CST), Danvers, MA, USA] for 1 h at room temperature protected from light. DAPI reagent (Solarbio, Beijing, China) was used for nuclei staining. Finally, specimens were observed under a fluorescence microscope (Leica).
Dapi reagent
Manufactured by Solarbio
Sourced in China
DAPI (4',6-diamidino-2-phenylindole) is a fluorescent dye that binds strongly to the minor groove of DNA. It is commonly used in fluorescence microscopy and flow cytometry applications to stain and visualize cell nuclei.
Automatically generated - may contain errors
Lab products found in correlation
Anti α1 ar antibody, by Abcam (1 mentions)
Fluorochrome conjugated secondary antibody, by Cell Signaling Technology (1 mentions)
15 mm glass bottom dishes, by NEST Biotechnology (1 mentions)
Fluorescence microscope, by Leica (1 mentions)
Edu reagent, by Keygen Biotech (1 mentions)
Tcs sp8, by Leica (1 mentions)
3 protocols using dapi reagent
1
Immunocytochemistry of α1-Adrenergic Receptor
2
Cell Proliferation Assay using EdU
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The DB or SU-DHL-4 cells were stained with 10 µM EdU reagent (Nanjing KeyGen Biotech. Co., Ltd.) at 37°C for 2 h, and fixed with 4% paraformaldehyde for 15 min. Subsequently, all cells were washed with PBS containing 3% bovine serum albumin (BSA), incubated with 0.5% Triton X-100 for 20 min and incubated with Click-iT reagent (Nanjing KeyGen Biotech. Co., Ltd.) at room temperature for 30 min in the dark. Finally, the cells were stained with DAPI reagent (Beijing Solarbio Science & Technology Co., Ltd.) at room temperature for 5 min, and photographed at a ×400 magnification.
3
Subcellular Localization of PfbZIP85 Protein
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To investigate the subcellular localization of PfbZIP85 protein, the recombinant fusion plasmid 35S::PfbZIP85::GFP (pCAMBIA1300-PfbZIP85/GFP) and the control plasmid 35S::GFP were transiently expressed in N. benthamiana leaf epidermal cells through Agrobacterium-mediated transformation [6 (link)]. After the tobacco plants were grown at 25 °C for more than 36 h, the transfected tobacco leaf samples were treated with DAPI reagent (Solarbio Life Sciences, Beijing, China), followed by observation of fluorescence signals utilizing a laser scanning confocal microscope (Leica TCS SP8).
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