4 6 diamidino 2 phenylindole c1002

Manufactured by Beyotime

Sourced in China

4',6-diamidino-2-phenylindole (C1002) is a fluorescent dye that binds to DNA. It is commonly used in biological research as a nuclear counterstain.

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Lab products found in correlation

Antifade mounting medium, by Abcam (1 mentions) A0562, by Beyotime (1 mentions) Lsm 880 confocal microscope, by Zeiss (1 mentions) Fluorescence microscope, by Leica (1 mentions) Zli 9056, by ZSGB-BIO (1 mentions) Confocal laser scanning microscope, by Leica (1 mentions) Ab115730, by Abcam (1 mentions) A0423, by Beyotime (1 mentions) Image pro plus 6, by Media Cybernetics (1 mentions) Bs6580, by Bioworld Technology (1 mentions) Ab52615, by Abcam (1 mentions) Bx43f, by Olympus (1 mentions) Cm3050, by Leica (1 mentions) P0126, by Beyotime (1 mentions) Eclipse ti sr, by Nikon (1 mentions)

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C1002 (104 citations) 4,6-diamidino-2-phenylindole (DAPI, C1002) (10 citations)

9 protocols using 4 6 diamidino 2 phenylindole c1002

Immunofluorescence Analysis of Renal AQP2 and Caspase-1

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The collected renal tissues of the mice were formalin-fixed, paraffin-embedded and then sectioned for subsequent haematoxylin-eosin (HE) staining. The following procedures were performed as previously described.
Immunofluorescence staining was performed to assess the expression of AQP2 and caspase-1. For frozen-section immunofluorescence staining, sections were fixed, blocked and then stained with the primary antibodies AQP2 (1:500, PAA580Hu01; Cloud-Clone Corp., Wuhan, China) and caspase-1 (1:100, PAB592Hu01; Cloud-Clone Corp.). The fluorescent secondary antibodies were fluorescein isothiocyanate-labeled goat anti-rabbit immunoglobulin G (1:500, A0562; Beyotime Biotechnology Co., Ltd., Haimen, Jiangsu, China) and 4′,6-diamidino-2-phenylindole (C1002; Beyotime Biotechnology Co., Ltd.). Sections were then mounted with anti-fade mounting medium (Abcam Co., Ltd, Cambridge, MA, USA).
FAM-FLICA kits (ICT098; ImmunoChemistry Technologies LLC., Bloomington, MN, USA) were utilized to conduct FAM-YVAD-FMK/Hoechst/propidium iodide (PI) staining according to the manufacturer’s instructions. Images were collected using a fluorescence microscope.

Fan Y., Ma M., Feng X., Song T., Wei Q, & Lin T. (2021). Overexpression of aquaporin 2 in renal tubular epithelial cells alleviates pyroptosis. Translational Andrology and Urology, 10(6), 2340-2350.

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Immunofluorescence Staining of Retinal Ganglion Cells

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The immunofluorescence staining was performed as previously described (Akopian et al., 2014). Mice were anesthetized as described above and sacrificed by cervical dislocation. After fixation in 4% paraformaldehyde for 30 minutes at 26°C, the eyes were immersed in 30% sucrose overnight and then embedded in optimal cutting temperature compound (Cat# 4583; Sakura, Oakland, CA, USA). The frozen specimens were sectioned at 10 µm thickness on a glass slide. Sections were blocked with 0.5% bovine serum albumin and 0.1% Triton X-100 for 1 hour at 26°C and incubated with primary antibodies overnight at 4°C. After washing, slides were incubated with secondary antibodies conjugated with Alexa Fluor 488/594 for 1 hour at 26°C and 4′,6-diamidino-2-phenylindole (C1002, Beyotime, Shanghai, China) at 26°C for 1 hour. All antibody information was shown in Table 1. After washing, slides were mounted in Vectashield media (P0126, Beyotime). Immunolabeled images were captured using an LSM 880 confocal microscope (Carl Zeiss, Jena, Germany). For RGC counting using anti-RNA-binding protein with multiple splicing (RBPMS), sections were selected across the optic nerve, and RBPMS-positive RGCs were counted from one ora serrata to the other.

Gan Y.J., Cao Y., Zhang Z.H., Zhang J., Chen G., Dong L.Q., Li T., Shen M.X., Qu J, & Chi Z.L. (2023). Srgap2 suppression ameliorates retinal ganglion cell degeneration in mice. Neural Regeneration Research, 18(10), 2307-2314.

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Quantifying Arecoline-Induced DNA Damage

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Immunofluorescence was applied to examine the expression of phosphor-histone H2A.X (γH2A.X) in HOKs treated with different concentrations of arecoline. After senescence induction, the cells were seeded into 24-wells culture plates at 1 ×10⁴ per well density. The next day, the cells were fixed with 4% paraformaldehyde for 10 min. Subsequently, the cells were permeabilized with 0.1% Triton X-100 solution and blocked with 10% goat serum (ZLI-9056, ZSGB-BIO). After PBS washing, the cells were incubated with monoclonal rabbit anti- γH2A.X primary antibody (1:400, 9718, Cell Signaling Technology) at 4° overnight. The next day, the cells were incubated with cy3-conjugated secondary antibody (AP132C, Sigma) for 1 h in dark, and then counterstained with 4′,6-Diamidino-2-phenylindole (C1002, Beyotime) at room temperature for 1 min. Immunofluorescence images were acquired using a fluorescence microscope (Leica Microsystems, Cambridge, UK). A total of five fields in each group were selected under the microscope and the total cells and γH2A.X positive cells in each field were counted by ImageJ 5.0. The proportion of γH2A.X positive cells was obtained by calculating the ratio of the average γH2A.X positive cell number to the average total cell number in each group.

Wang Z., Han Y., Peng Y., Shao S., Nie H., Xia K., Xiong H, & Su T. (2023). Senescent epithelial cells remodel the microenvironment for the progression of oral submucous fibrosis through secreting TGF-β1. PeerJ, 11, e15158.

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Visualizing Cytoskeletal Proteins

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Confocal fluorescence microscopy was used for analyzing expressions of Vimentin and α-smooth muscle actin (α-SMA). The cells were washed with ice PBS and incubated in specific primary antibodies (anti-vimentin (bs-23063R) and anti-α-SMA (bs-10196R), Bioss Inc. Beijing, China). Then cells were washed again and incubated in fluorescein isothiocyanate-labeled anti-rabbit immunoglobulin G antibody (A0562, Beyotime, China) and 4′,6-diamidino-2-phenylindole (C1002, Beyotime, China). Vimentin and α-SMA that were fluorescent labeled could be visualized by the confocal microscope.

Hou L., Le G., Lin Z., Qian G., Gan F., Gu C., Jiang S., Mu J., Ge L, & Huang K. (2020). Nontoxic concentration of ochratoxin A decreases the dosage of cyclosporine A to induce chronic nephropathy model via autophagy mediated by toll-like receptor 4. Cell Death & Disease, 11(2), 153.

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Visualizing Drug Localization in Cells

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SNU16, K562 and HGC27 cells were treated with JQ1 (2 µM), JQ1-btn (2 µM), THZ1 (1 µM), THZ1-btn (1 µM), Dox (3 µM) or Dox-btn (3 µM) for 3 h, respectively. Cells were fixed with 0.25% formaldehyde for 5 min on ice. CRC organoids were treated with JQ1 (5 µM), JQ1-btn (5 µM), THZ1 (5 µM), THZ1-btn (5 µM), Dox (3 µM) or Dox-btn (3 µM) for 6 h. Subsequently, the organoids were fixed with 4% formaldehyde for 10 min. After washing twice with 0.1% BSA/PBS, cells were dropped onto the slides for 2 h at room temperature. Slides were then permeabilized with 0.5% TX-100 in PBS and blocked with 3% BSA/PBS for 30 min. Finally, slides were incubated overnight at 4 °C with primary rabbit monoclonal anti-biotin (CST, 5597S, 1:250 dilution), and goat anti-rabbit Alexa Fluor 488 secondary antibodies (4412S, CST) were used for visualization of drug. Cell nuclei were stained with 4,6-diamidino-2-phenylindole (C1002, Beyotime), and the slides were scanned under a confocal microscope.

Dong C., Meng X., Zhang T., Guo Z., Liu Y., Wu P., Chen S., Zhou F., Ma Y., Xiong H., Shu S, & He A. (2024). Single-cell EpiChem jointly measures drug-chromatin binding and multimodal epigenome. Nature methods.

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Quantitative Analysis of Glucose Transporter 1

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After being fixed and permeabilized, cells were fixed, permeabilized, blocked, and incubated with anti-GLUT1 (1:500; ab115730; Abcam) and Alexa Fluor 488-labeled Goat Anti-Rabbit IgG (H + L) (1:500; A0423; Beyotime Biotechnology) antibodies. 4',6-diamidino-2-phenylindole (C1002; Beyotime Biotechnology) labeling was used to visualize cell nuclei. Then, visualization of the positively stained cells was performed using confocal laser scanning microscope (Leica Microsystems, Inc., Deerfield, IL, USA).

Tan M., Pan Q., Yu C., Zhai X., Gu J., Tao L, & Xu D. (2024). PIGT promotes cell growth, glycolysis, and metastasis in bladder cancer by modulating GLUT1 glycosylation and membrane trafficking. Journal of Translational Medicine, 22, 5.

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Immunofluorescence Staining of CHOP, IRE1α, Ar, Cyp19α1, and Cyp11α1

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Immunofluorescence was conducted as described previously for immunohistochemistry with slight modification. Essentially, after incubation with the primary antibody against CHOP (1 : 200), IRE1α (1 : 50), p-IRE1α, Ar (1 : 200, ab52615, Abcam, Cambridge, UK), Cyp19α1 (1 : 200, BS6580, Bioworld, Minnesota, USA), and Cyp11α1 (1 : 200, BS5680, Bioworld) overnight at 4°C, tissue sections and cells were incubated at 24°C for 2 h with fluorescently labeled secondary antibodies. Nuclei were counterstained with 4′,6-diamidino-2-phenylindole (C1002, Beyotime, Shanghai, China) at a dilution of 1 : 2000 for 30 min. Images were photographed using an Olympus laser scanning confocal microscope (FV3000). Fluorescence intensity was quantified using Image-Pro Plus 6.0 (Media Cybernetics, Maryland, USA).

Zhang Y., Weng Y., Wang D., Wang R., Wang L., Zhou J., Shen S., Wang H, & Wang Y. (2021). Curcumin in Combination with Aerobic Exercise Improves Follicular Dysfunction via Inhibition of the Hyperandrogen-Induced IRE1α/XBP1 Endoplasmic Reticulum Stress Pathway in PCOS-Like Rats. Oxidative Medicine and Cellular Longevity, 2021, 7382900.

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Immunofluorescence Analysis of BACE1, PS1, and ADAM10

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After treatment with different concentrations of ICT for 16 h, cells plated on coverslips were fixed with methanol for 20 min at 4 °C. They were permeabilized with 0.1% Triton X-100 in phosphate-buffered saline for 5 min at room temperature, and treated with blocking medium (5% bovine serum albumin in phosphate-buffered saline) for 30 min. Then, they were incubated with antibodies against anti-BACE1 (1:1,000 dilution), anti-PS1 (1:1,000), and anti-ADAM10 (1:1,000) at room temperature for 2 h. Immune-reacted primary antibody was detected following 1 h incubation in the dark at room temperature with a secondary antibody conjugated with Dylight 488 (1:1,000 dilution). Cells were further stained with 4′, 6-diamidino-2-phenylindole (C1002; Beyotime) for 2 min in the dark at room temperature and washed. Then, they were placed onto microscope slides in mounting medium. Observations were carried out using a fluorescence microscope (BX43F; Olympus, Tokyo, Japan).

Feng F., Li Y., Huang N, & Luo Y. (2019). Icaritin, an inhibitor of beta-site amyloid cleaving enzyme-1, inhibits secretion of amyloid precursor protein in APP-PS1-HEK293 cells by impeding the amyloidogenic pathway. PeerJ, 7, e8219.

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TUNEL Analysis of Apoptosis in Mouse Brains

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Mouse brains were removed after treatments with or without sevoflurane and stored at 4°C in 4% paraformaldehyde. Serial coronal sections (10 µm) were cut on a cryostat (CM3050 S, Leica Biosystems, Germany) and mounted on coverslips. Sections were permeabilized with proteinase K solution (20 µg/mL) for 20 min. Terminal deoxynucleotidyl transferase (TdT) and dUTP (11684817910, Roche) were added to the sections and incubated in a humidified chamber at 37°C for 2 h. The reaction was then stopped and the sections were stained with 4'6'-diamidino-2phenylindole (C1002, Beyotime, China) for 10 min. Then, coverslips were mounted on glass slides with anti-fade mounting medium (P0126, Beyotime, China). Finally, the sections were analyzed under a light microscope (ECLIPSE TI-SR, NIKON, Japan) with 5X and 20X objective lenses, and photographs of the sections were taken. The number of TdT dUTP nick-end labeling (TUNEL)positive cells was counted using the Image J software by an investigator who was blinded to the experimental design.

Song Q., Ma Y.L., Song J.Q., Chen Q., Xia G.S., Ma J.Y., Feng F., Fei X.J, & Wang Q.M. (2015). Sevoflurane induces neurotoxicity in young mice through FAS/FASL signaling. Genetics and molecular research : GMR, 14(4).

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