Paraffin-embedded kidney tissue sections were deparaffinized and subjected to antigen retrieval, which was performed under high pressure in citrate buffer (0.01 mol/L, pH 6.0) for 10 min. After blocking with 5% bovine serum albumin (BSA) for 1 h, the sections were incubated with primary antibodies overnight at 4°C (Nrf2, GeneTex, USA, #GTX103322; synaptopodin, Progen, Germany, #65194), followed by incubation with fluorochrome-conjugated secondary antibodies (Thermo Fisher Scientific) at 37°C for 2 h in the dark. Nuclei were counterstained with 4′,6′-diamidino-2-phenylindole (DAPI, Antgene, China). All images were taken with a fluorescence microscope (Olympus, Japan).
4 6 diamidino 2 phenylindole dapi
Manufactured by Antgene
Sourced in China
4',6'-diamidino-2-phenylindole (DAPI) is a fluorescent dye that binds strongly to adenine-thymine (A-T) rich regions in DNA. It is commonly used in various biological applications as a nuclear counterstain.
Automatically generated - may contain errors
Lab products found in correlation
Synaptopodin, by Progen Biotechnik (1 mentions)
Fluorochrome conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions)
Fluorescence microscope, by Olympus (1 mentions)
Donkey serum, by Antgene (1 mentions)
Desmin, by R&D Systems (1 mentions)
α sma, by R&D Systems (1 mentions)
Cd151, by Abcam (1 mentions)
Eight well chamber slide, by Ibidi (1 mentions)
Image pro plus, by Olympus (1 mentions)
3 protocols using 4 6 diamidino 2 phenylindole dapi
1
Immunofluorescence Analysis of Kidney Tissue
2
Immunostaining of Cell-climbing Films
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The cell-climbing films were fixed with 4% paraformaldehyde for 30 min at 4 °C and incubated first with a guinea pig anti-ADRP antibody (1:100, PROGEN Biotechnik, Germany) overnight at 4 °C, and then with tetramethylrhodamine (TRITC)-conjugated goat anti-guinea pig immunoglobulin G (IgG) (1:100, Thermo Scientific, USA) at 37 °C for 60 min in the dark. The nuclei were counterstained with 4′, 6-diamidino-2-phenylindole (DAPI) (Antgene, China) for 5 min. All microscopy images were recorded using a confocal microscope (Olympus, Japan).
3
Characterization of Mesenchymal Stem Cells
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Gb-MSCs and bm-MSCs were grown on an eight-well chamber slide (Ibidi, Germany) until 70% confluent and washed with PBS three times. Then, the medium was replaced with serum-free medium (0% DMEM) and serum-free glioblastoma-conditioned medium (0% gb-CM), and the cells were incubated for 72 hours at 37°C. The cells were fixed with 4% paraformaldehyde, permeabilized with Triton-X-100 and blocked with solution containing donkey serum (Antgene, WuHan; China) for 1 h at room temperature. Then, the cells were incubated with primary antibodies against α-SMA, Desmin (RD, USA), NG2, Nestin, CD31, CD151, VE-cadherin (VE-cad) or smooth muscle myosin (SMM) (Abcam, USA) overnight at 4°C. After being washed with PBS, the cells were incubated with secondary antibodies labeled with either fluorescein or rhodamine (Antgene, WuHan; China) for 1 h at room temperature. In each experiment, nonspecific staining by secondary antibodies was excluded by incubating a well without primary antibodies. The staining was visualized using 4', 6-diamidino-2-phenylindole (DAPI) (Antgene, WuHan; China). A digital image was acquired using an Olympus (Olympus, Japan) camera, and the results were analyzed using Image-Pro Plus.
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