Product overview

Name DAPI
Description

Blue fluorescent DNA stain. Nuclear counterstain. Also available in solution.

Biological description

Overview

DAPI is a blue fluorescent DNA stain which is cell permeant at high concentrations.

DAPI binds strongly to A-T rich regions in DNA to form a fluorescent complex. It preferentially stains ds-DNA and has a high quantum yield (φf=0.92) when bound to DNA.

Uses and applications

DAPI is commonly used as a nuclear and chromosome counterstain.

It is preferentially used to stain dead cells. DAPI is less effective as a live cell stain as it is unable to efficiently pass through the membrane in live cells. Therefore, higher concentrations may need to be used.

Cells must be permeabilized and/or fixed for DAPI to enter the cell and bind to DNA.

Due to DAPI’s blue emission, there is very little fluorescent overlap between yellow-fluorescent, green-florescent molecules (e,g, fluorescein and GFP) or red-fluorescent stains (e.g. Texas red). It is therefore convenient for multiplexing assays.

DAPI has a great variety of applications but is often used for cell imaging, cell counting, cell sorting (based on DNA content), apoptosis analysis and in HCA (high-content analysis).

 

DAPI Staining Solution (1mg/mL) also available.

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Images

Fig 1: Neurofilament L and DAPI co-staining in hippocampal cell culture.
Fig 2: GFAP and DAPI co-staining in hippocampal cell culture.
Fig 3: MAP2 and DAPI co-staining in hippocampal cell culture.
Fig 4. Peak emission spectrum of DAPI when excited at 360nm and excitation spectrum when recording at 460nm

Biological Data

Application notes

Figure 1: Neurofilament L and DAPI co-staining in hippocampal cell culture.

DAPI is a DNA binding dye commonly used to label cell nuclei in immunofluorescence experiments. DAPI from Hello Bio labels cell nuclei (blue) at 1µg/ml when co-stained with an anti-neurofilament L antibody (green). For protocol see #Protocol 1 in application notes below.

 

Figure 2: GFAP and DAPI co-staining in hippocampal cell culture.

DAPI is a DNA binding dye commonly used to label cell nuclei in immunofluorescence experiments. DAPI from Hello Bio labels cell nuclei (blue) at 1µg/ml when co-stained with an anti-GFAP antibody (green). For protocol see #Protocol 1 in application notes below.

 

Figure 3: MAP2 and DAPI co-staining in hippocampal cell culture.

DAPI is a DNA binding dye commonly used to label cell nuclei in immunofluorescence experiments. DAPI from Hello Bio labels cell nuclei (blue) at 1µg/ml when co-stained with an anti-MAP2 antibody (green). For protocol see #Protocol 1 in application notes below.

 

Figure 4. Peak emission spectrum of oxazole yellow when excited at 360nm and excitation spectrum when recording at 460nm.

DAPI is a DNA binding dye with strong affinity for nucleic acids used to label cell nuclei in immunofluorescence experiments. DAPI from Hello Bio has an excitation peak at 361nm and an emission peak at 456nm. For protocol see #Protocol 2 in application notes below.

 

#Protocol 1: DAPI counterstaining of primary cultured neurones.

  • Primary neurones were isolated and cultured from P2 rats and grown for three weeks before being fixed with 4% paraformaldehyde.
  • Coverslips containing neuronal cell cultures were labelled for either MAP2, GFAP or Neurofilament L following standard immunohistochemical approaches.
  • Coverslips were then submerged in 1µg/ml DAPI diluted in PBS for 1 minute.
  • Following 2 x 5-minute washes in PBS coverslips were mounted and imaged with a fluorescent microscope.

 

#Protocol 2: Measurement of excitation and emission spectra of DAPI

  •  DAPI was prepared at 10 ng/ml in PBS with 10 ng/ml empty plasmid DNA.
  • Excitation and emission spectra were measured between 230nm and 700nm using a Tecan Infinite M200 PRO ELISA plate reader.

 

Solubility & Handling

Storage instructions -20°C
Solubility overview Soluble in water (10mg/ml, gentle warming), and in methanol
Important This product is for RESEARCH USE ONLY and is not intended for therapeutic or diagnostic use. Not for human or veterinary use.

Calculators

Molarity

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Dilution

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Chemical Data

Chemical name 4',6-Diamidino-2-phenylindole dihydrochloride
Molecular Weight 350.24
Chemical structure Chemical structure
Molecular Formula C16H15N5.2HCl
CAS Number 28718-90-3
PubChem identifier 160166
SMILES C1=CC(=CC=C1C2=CC3=C(N2)C=C(C=C3)C(=N)N)C(=N)N.Cl.Cl
InChiKey FPNZBYLXNYPRLR-UHFFFAOYSA-N
MDL number MFCD00012681
Excitation 340 / 360nM (for ds-DNA)
Emission 488 / 460nM (for ds-DNA)

References for DAPI

References are publications that support the biological activity of the product
  • The use of DAPI fluorescence lifetime imaging for investigating chromatin condensation in human chromosomes.

    Estandarte et al (2016) Sci Rep. 16 : 6-31417
  • Analysis of Apoptosis and Necroptosis by Fluorescence-Activated Cell Sorting.

    Wallberg et al (2016) Cold Spring Harb Protoc 4 : 087387
  • New insights into the in situ microscopic visualization and quantification of inorganic polyphosphate stores by 4',6-diamidino-2-phenylindole (DAPI)-staining.

    Gomes FM et al (2013) Eur J Histochem 57(4) : e34.
  • Labeling nuclear DNA using DAPI.

    Chazotte B (2011) Cold Spring Harb Protoc 2011(1) : pdb.prot5556.
  • Labeling nuclear DNA using DAPI.

    Chazotte et al (2011) Cold Spring Harb Protoc 2011(1) : 5556