DAPI

(HB0747)
Technical documents: SDS Datasheet

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

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.

 

#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.

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 DAPI  [28718-90-3] 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
Publications
These publications cite the use of DAPI purchased from Hello Bio:
  • No evidence from complementary data sources of a direct projection from the mouse anterior cingulate cortex to the hippocampal formation

    Craig et al (2022) Biorxiv : https://doi.org/10.1101/2022.01.25.477805

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