DAPI Staining Solution (Ready-to-use)

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  Immunofluorescent analysis of (4% PFA) fixed SKOV-3 cells using Alpha Tubulin antibody (66031-1-Ig, Clone: 1E4C11 ) at dilution of 1:1000 and CoraLite®594-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L), Nucleus was labelled in blue with DAPI.

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  Immunofluorescent analysis of (4% PFA) fixed paraffin-embedded mouse testis tissue using DAPI. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  Immunofluorescent analysis of (4% PFA) fixed paraffin-embedded mouse spleen tissue using DAPI. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  Immunofluorescent analysis of (4% PFA) fixed HepG2 cells using DAPI.

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  Immunofluorescent analysis of (4% PFA) fixed frozen OCT-embedded mouse brain tissue using DAPI (PR30021).

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  Immunofluorescent analysis of (4% PFA) fixed frozen OCT-embedded rat brain tissue using DAPI (PR30021).

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  2.5x10^6 HeLa cells were intracellularly stained with DAPI (PR30021, blue). Cells were fixed with 4% PFA.

Cat No. PR30021

Publications

6

CAS No.

28718-90-3

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Product Information

DAPI, also known as DAPI dihydrochloride (4',6-diamidino-2-phenylindole), is a commonly used blue fluorescent nucleic acid stain. It binds to the minor groove of AT-rich base pair sequences in double-stranded DNA, and its blue fluorescence is enhanced more than 20-fold upon binding to DNA. It is frequently used for nuclear staining of fixed cells and tissue sections, which can be observed under a fluorescence microscope after staining.

When DAPI binds to DNA, its maximum excitation wavelength is 358 nm, and its maximum emission wavelength is 461 nm. Although DAPI is not membrane-permeable to live cells under normal conditions, it can enter live cells at elevated concentrations. Upon excitation, DAPI emits blue fluorescence, allowing it to be combined with other fluorochromes such as green, orange, and red ones to achieve multiplex fluorescent labeling.

This product is a ready-to-use DAPI aqueous solution, which can be directly applied to nuclear immunofluorescence staining of fixed cells or tissue sections, as well as flow cytometry.

Product Name	DAPI Staining Solution (Ready-to-use)
Alternative Name	4',6-diamidino-2-phenylindole
Molecular Formula	C16H15N5·2HCl
Molecular Weight	350.25
CAS No.	28718-90-3

Storage

Store at 2–8°C away from light. Valid for one year from the date of receipt.

Usage

Immunofluorescence Staining:

1. Staining: If other fluorescent staining is to be performed, immunofluorescence staining may be conducted first, followed by DAPI staining as the final step. If no other fluorescent staining is required, DAPI staining can be performed directly.

2. Add an appropriate volume of DAPI working solution to fixed adherent cells or tissue sections to fully immerse the samples, and incubate at room temperature for 8-10 minutes. For fixed suspension cells, add at least 3 times the volume of the staining solution relative to the sample volume, and mix thoroughly.

3. Stain at room temperature for 8-10 minutes.

4. Aspirate the DAPI staining solution, and wash with PBS 2-3 times, for 3-5 minutes each time.

5. Observe under a fluorescence microscope at Ex/Em = 358 nm/461 nm.

Flow Cytometry:

1. Collect the cultured cells into EP tubes, centrifuge at 300-400 × g for 5 minutes at room temperature, and discard the supernatant. Add 1 mL of PBS and pipette gently to ensure complete cell resuspension, and repeat this process three times.

2. Add 0.5 mL of 4% PFA or absolute ethanol to each EP tube containing 1 × 10⁶ cells, fix at room temperature for 15 minutes, centrifuge at 400–600 × g for 5 minutes at room temperature, and discard the supernatant.

3. Add 1 mL of PBS to the EP tube, pipette gently to ensure complete cell resuspension, centrifuge at 400–600 × g for 5 minutes, and discard the supernatant.

4. Repeat Step 3 three times.

5. Add 0.2 mL of DAPI working solution to each EP tube containing 1 × 10⁶ cells, and pipette gently to ensure complete cell resuspension. Incubate at room temperature for 10-15 minutes, protected from light.

6. Analyze the samples on a flow cytometer.

Notes

1. DAPI is irritating to humans. Proper protective measures must be taken during operation. Please wear a lab coat and disposable gloves to avoid direct skin contact and cross-contamination.

2. All fluorescent dyes are subject to fluorescence quenching. It is recommended that detection be completed on the same day after staining. Antifade reagents can be used to mitigate fluorescence quenching.

3. This product should be stored and used away from light, and repeated freeze-thaw cycles should be avoided.

4. This product is for research use only.

Cited in Article as

PR30021, DAPI Staining Solution (Ready-to-use), Proteintech, IL, USA

Publications

Application	Title
	Front Biosci (Landmark Ed); Co-Highly Expressed SLC17A9 and KCNH1 as Potential Prognostic Biomarkers and Therapeutic Targets in Clear Cell Renal Cell Carcinoma; Authors - Zongpan Ke
IF	Adv Healthc Mater; Multifunctional Sr2+/Zn2+ Co-Doped Mesoporous Silica Nanoparticles in Injectable Hydrogel for Ameliorating Osteoporotic Osseointegration; Authors - Wei Cui
	Cell Mol Life Sci; Progranulin enhances M2 macrophage polarization and renal fibrosis by modulating autophagy in chronic kidney disease; Authors - Wei-Chao Tu
	Neural Regen Res; Changes in border-associated macrophages after stroke: single-cell sequencing analysis; Authors - Ning Yu
IF	Int Immunopharmacol; H₂O₂ preconditioning enhances the transplantation-mediated therapeutic effect of bone marrow-derived mesenchymal stem cells after traumatic brain injury; Authors - En-Yan Jiang
IF	Life Sci; SETD1B promotes brain cell ferroptosis in ischemic stroke mice via increasing H3K4me3 enrichment on the Tfrc promoter; Authors - Li Wang