5 μm frozen sections of kidney specimens were washed three times with PBS. Then slides were blocked with 5% bull serum albumin (BSA) for half an hour and incubated with both TRPM2 antibody and the antibody of specific renal tubular epithelial cell marker, antiaquaporin 1 (AQP1) overnight at 4° C (Supplementary Table 1 ). After washing with PBS, slides were correspondingly incubated with two kinds of secondary antibody (Cy3 conjugate, SA00009-1, Proteintech, Wuhan, 1:100; FITC conjugate SA00003-1, Proteintech, Wuhan, 1:100, respectively) for 1 h at room temperature in the darkroom. The cell nuclei were stained with 4',6-diamidino-2-phenylindole (DAPI, C0065, Solarbio, Beijing) for 10 min, and all stained sections were viewed by fluorescent microscope (Olympus, Tokyo).
4 6 diamidino 2 phenylindole dapi c0065
Manufactured by Solarbio
Sourced in China
4',6-diamidino-2-phenylindole (DAPI, C0065) is a fluorescent stain that binds strongly to adenine-thymine (A-T) rich regions in DNA. It can be used to visualize and quantify DNA in various applications.
Automatically generated - may contain errors
Lab products found in correlation
Lipofectamine rnaimax, by Thermo Fisher Scientific (2 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions)
Trizol reagent, by Thermo Fisher Scientific (2 mentions)
Fluorescent microscope, by Olympus (1 mentions)
Anti ubiquitin p4d1 sc 8017, by Santa Cruz Biotechnology (1 mentions)
Anti β actin ac038, by ABclonal (1 mentions)
Horseradish peroxidase hrp conjugated secondary goat anti rabbit ak1007 and goat anti mouse ak801x antibodies, by Sungene Biotech (1 mentions)
Chromotek gfp trap, by Proteintech (1 mentions)
Pp2 hy 13805, by MedChemExpress (1 mentions)
3 protocols using 4 6 diamidino 2 phenylindole dapi c0065
1
Immunofluorescence Staining of Renal Tubules
2
Regulation of HIF-1α and O-GlcNAcylation
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The antibodies against O-linked N-acetylglucosamine (O-GlcNAc) (RL2, ab2739) were purchased from Abcam. Anti-OGT (F-12,sc-74546), anti- O-GlcNAcase (OGA) (G-12,sc-376429), anti–HIF–1α (28b,sc-13515), as well as anti-ubiquitin (P4D1，sc-8017) were obtained from Santa Cruz Biotechnology. Anti-fibronectin (A12932), anti-COL1A2 (A5786), anti-E-Cadherin (A11509), anti-α-SMA (A17910), as well as anti-β-Actin (AC038) were purchased from Abclonal (China). Horseradish peroxidase (HRP)-conjugated secondary goat anti-rabbit (AK1007) and goat anti-mouse (AK801X) antibodies, as well as TRITC Goat anti-mouse antibody (GM200G-39C) were purchased from SUNGENE BIOTECH (China). The 4′-6-diamidino-2-phenylindole (DAPI) (C0065) was obtained from Solarbio (China) and PUGNAc (ab144670) was obtained from Abcam. OGT inhibitor OSMI-1 (T16409), MG-132 (T2154) and FG-4592 (T2515) were obtained from TargetMol. HIF-1α inhibitor (sc-205346) was purchased from Santa Cruz Biotechnology.
3
DAPI-Based Cellular Labeling Protocol
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4′,6-Diamidino-2-phenyl-indole (DAPI) (C0065) was purchased from Solarbio (Beijing, China). ChromoTek GFP-Trap (gta) and DYKDDDDK Fab-Trap (ffa) were purchased from Proteintech (Wuhan, China). BeaverBeads GSH (70601-K10) and His tag (70501-K10) were purchased from Beaverbio (Suzhou, China). TRIzol reagent (15596018), Lipofectamine 2000 (11668019), and Lipofectamine RNAiMAX (13778150) were purchased from Thermo Fisher Scientific (Waltham, MA, USA). A2ti-1 (HY-136465) and PP2 (HY-13805) were purchased from MedChemExpress (South Brunswick, NJ, USA).
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