4 6 diamidino 2 phenylindole (dapi)

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About the product

DAPI is a fluorescent dye that binds strongly to adenine-thymine (A-T) rich regions in DNA. It is commonly used as a nuclear counterstain in fluorescence microscopy to visualize and locate cell nuclei.

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Market Availability & Pricing

4',6-Diamidino-2-phenylindole (DAPI) is an actively commercialized product by the Merck Group under its Sigma-Aldrich brand. The product is available for purchase through the official Sigma-Aldrich website.

Prices for DAPI vary depending on the quantity and packaging. A 5 mg package is available at a price range of approximately £90 to $110.

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Spelling variants (same manufacturer)

Diamidino-2-phenylindole (DAPI) - 73 citations 4′-DAPI - 5 citations

The spelling variants listed below correspond to different ways the product may be referred to in scientific literature.
These variants have been automatically detected by our extraction engine, which groups similar formulations based on semantic similarity.

22 622 protocols using «4 6 diamidino 2 phenylindole (dapi)»

Organoid Immunofluorescence Staining Protocol

2025

Immunofluorescence staining experiments were performed on organoids as previously described^{98 (link)}. When the organoids reached a size of ~100 μm, they were selected for staining. After washing twice with pre-cooled DPBS, 500 μL of cell recovery solution (354253, Corning) was added to each well, and the Matrigel was dissolved on ice to ensure that the morphology of the organoids was not disrupted. After 30 min, all the organoids were collected into a 15 mL centrifuge tube, fixed with 4% paraformaldehyde for 30 min, and then centrifuged to remove the supernatant. Next, 10 mL of 1% PBST was added to stop the tissue fixation. After blocking with Organoid Washing Buffer (OWB, 0.1% Triton X-100, 0.2% BSA in DPBS), the primary antibody was added and incubated overnight at 4 °C with shaking at 60 rpm. On the following day, the organoids were washed three times with OWB for 2 h each time, and then the corresponding fluorescent secondary antibody was added. The organoids were incubated overnight on a shaker in the dark. On the third day, 4′,6-Diamidino-2-phenylindole dihydrochloride (DAPI, D9542, Sigma) at 10 mg/mL was added for 30 min. After washing with OWB, the samples were spun down at 70 × g for 5 min at 4 °C. Finally, the organoids were resuspended with fructose-glycerol clearing solution (60% glycerol and 2.5 M fructose in ddH₂O) and imaged using an LSM880 confocal microscope (Zeiss) and a CSU-W1 spinning disk field scanning confocal system (Nikon). A cell death detection (TUNEL) kit (Roche) was used to identify dead cells in accordance with the company’s description. All the antibodies used in this study were listed in Supplementary Data 10.

Zhang N., Meng Y., Mao S., Ni H., Huang C., Shen L., Fu K., Lv L., Yu C., Meekrathok P., Kuang C., Chen F., Zhang Y, & Yuan K. (2025). FBXO31-mediated ubiquitination of OGT maintains O-GlcNAcylation homeostasis to restrain endometrial malignancy. Nature Communications, 16, 1274.

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Immunofluorescence Staining Protocol

2025

Cells were fixed with 4% paraformaldehyde for 10 min and washed in PBS. Cells were permeabilized (PBS 1×, 20 mM Glycine, 0.1% Triton) for 10 min and then blocked (PBS 1×, 20 mM Glycine, 1% BSA, 0.01% Triton) for 1 h. Next, cells were incubated with primary antibodies for 1 h and washed twice with washing solution (PBS 1×, 20 mM Glycine). Cells were then incubated with secondary antibodies for 45 min, followed by two washes. Coverslips were mounted using Vectashield mounting medium containing DAPI (Vector Laboratories) and Ibidi slides were left in PBS after being DAPI stained (Sigma‐Aldrich) for 10 min. All the incubations were performed at room temperature.

Bosch‐Calvet M., Pérez‐Venteo A., Cebria‐Xart A., Garcia‐Cajide M, & Mauvezin C. (2025). Nuclear stiffness through lamin A/C overexpression differentially modulates chromosomal instability biomarkers. Biology of the Cell, 117(2), e12001.

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Cell Cycle Analysis of Hematopoietic Stem and Progenitor Cells

2025

Single cell suspension from P0 neonatal liver and P14 BM were first stained with lineage markers (Ter119, Gr1, B220, CD3, CD4, CD5, CD8) and HSPC markers (cKit, Sca1, CD150, Flk2), then fixed with 4% PFA, permeabilized with 0.3% saponin, treated with 100U RNAase A (ThermoFisher), and stained with 0.01 mg/ml DAPI (Millipore) or Hoechst 33342 (ThermoFisher) for cell cycle analysis by flow cytometry.

Cool T., Baena A.R., Rommel M., Mattingly C., Bachinsky E., Saini S., Chattopadhyaya S., Manso B., Rajendiran S., Worthington A., Poscablo D., Deguzman A., Berger-Cahn T., Boyd D, & Forsberg E. (2025). Perinatal Nicotine Exposure Disrupts Hematopoietic Stem Cell Development and Elevates Influenza Susceptibility in Adulthood. bioRxiv.

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Flow Cytometry Immunophenotyping of Cells

2025

Cells were stained with 100 ng/mL DAPI (Sigma Aldrich, St Louis, MO) to discriminate living cells, treated with FcR blocking reagent (Miltenyi Biotec, Bergisch Gladbach, Germany), and then incubated with the specific conjugated antibodies listed in Table 3. Not-labelled cells were used to set the autofluorescence.Table 3

List of antibodies used in flow cytometry

Anti-human antibody	Fluorochrome	Brand	Dilution
CD45	APC	Miltenyi Biotec	1:50
CD34	PE	BD	1:25
CD43	FITC	BD	1:50
HLA-DR	PerCP-Cy5.5	BD	1:10
CD14	FITC	BioLegend	1:25
CD11b	APC	Miltenyi Biotec	1:100
CD16	PE	BD	1:10
CD49d	PE	eBioscience	1:20

The acquisition was performed on FACSCanto™ II (BD, Franklin Lakes, NJ) from at least 10⁴ events/tube. The data were elaborated with FlowJo software (TreeStar, Ashland, OR). The results are expressed either as percentage or as the difference between the mean fluorescence intensity of labelled cells and of the corresponding not-labelled control (ΔMFI), normalized on the ΔMFI of NT samples, according to the following formula:

Δ MFI ratio = \frac{{({MFI}_{labelled} - {MFI}_{unlabelled})}_{treat}}{{({MFI}_{labelled} - {MFI}_{unlabelled})}_{NT}}

Mazzoldi E.L., Benini G., Ferraro R.M., Micheletti M., Martellosio G., Balduchelli V., Sacristani P., Lussignoli D., Semeraro F., Rezzola S., Presta M., Bergandi L., Meduri A, & Giliani S.C. (2025). Differentiation of human hyalocytes from induced pluripotent stem cells through ascorbic acid treatment. Human Cell, 38(2), 52.

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Immunohistochemical Analysis of Spinal Cord and DRG

2025

Mice were euthanized with an injection of pentobarbital-phenytoin solution (i.p.) and transcardially perfused with 10 mL of ice-cold 0.9 % saline, followed by 10 mL of ice-cold 4 % paraformaldehyde. L4/5 spinal cord and L4/5 DRG were post-fixed in 4 % paraformaldehyde overnight prior to being cryoprotected stepwise in 15 %, 22.5 % and 30 % sucrose in 0.1 M phosphate buffer (PB) with 0.01 % sodium azide (Millipore Sigma, CA USA) at 4 °C. Tissues were freeze-mounted in optimal cutting temperature compound (Sakura Finetek, CA USA), and serially sectioned (12 μm for DRGs and 16 μm for spinal cords) using a cryostat (Leica Biosystems) and mounted to SuperFrost Plus charged slides (Thermo Fisher Scientific), which were stored at −20 °C. For DRG, slides were washed three times in 0.3 % Triton X-100 (Millipore Sigma) in PBS (PBST) and incubated with blocking buffer (5 % normal donkey serum (Abcam, MA, USA), in PBST) for 1hr. For spinal cord, sections were permeabilized in 0.1 % NaBH4 in PBS for 20 mins and washed twice (PBST) prior to blocking. Primary antibodies were added after washing and incubated at 4 °C overnight. Subsequently, sections were washed three times, then incubated with secondary antibodies for 2 hrs. Slides were then washed three times, incubated with DAPI (1:5,000; Sigma-Aldrich, MO USA) for 5 mins, washed in PBS four times, and cover slipped with FluorSave mounting medium (Millipore Sigma). Primary antibodies used were goat anti-mouse IgG antibody (1:100; Jackson ImmunoResearch, PA USA), rabbit anti-mouse IgG antibody (1:100; Jackson ImmunoResearch), rabbit anti-IBA1 (1:500, Wako, Japan), rat anti-F4/80 (1:1000, eBioscience, CA USA), rabbit anti-MAP2 (1:250, Abcam), goat anti-CD31 (1:250, R&D Systems), mouse anti-FcRγ (1:200, MBL Life science, Japan), mouse anti-FcRn (1:200, Santa Cruz Biotechnology, CA USA) and rabbit anti-NK.1 (1:100, Invitrogen). Secondary antibodies used were donkey anti-goat antibody, donkey anti-rabbit antibody, donkey anti-rat and donkey anti-mouse (1:500; Thermo Fisher Scientific). A list of primary antibodies and secondary antibodies is provided in Table S1.

Fiore N.T., Willcox K.F., Dayani D., Zuberi Y.A., Heijnen C.J, & Grace P.M. (2025). Reducing IgG accumulation via neonatal Fc receptor (FcRn) blockade relieves neuropathic pain. Brain, behavior, and immunity, 125, 371-387.

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