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D9542

Sigma-Aldrich

DAPI

for nucleic acid staining

Synonym(s):

4′,6-Diamidino-2-phenylindole dihydrochloride, 2-(4-Amidinophenyl)-6-indolecarbamidine dihydrochloride, DAPI dihydrochloride

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About This Item

Empirical Formula (Hill Notation):
C16H15N5 · 2HCl
CAS Number:
Molecular Weight:
350.25
Beilstein:
4894417
EC Number:
MDL number:
UNSPSC Code:
12352200
PubChem Substance ID:
NACRES:
NA.52

grade

for molecular biology

Quality Level

Assay

≥98% (HPLC)

form

powder

technique(s)

transfection: suitable

solubility

H2O: 20 mg/mL
PBS: insoluble

ε (extinction coefficient)

30 at 263 nm in H2O at 1 mM

fluorescence

λex 340 nm; λem 488 nm (nur DAPI)
λex 364 nm; λem 454 nm (DAPI-DNA-Komplex)

suitability

suitable for fluorescence

storage temp.

2-8°C

SMILES string

Cl.Cl.NC(=N)c1ccc(cc1)-c2cc3ccc(cc3[nH]2)C(N)=N

InChI

1S/C16H15N5.2ClH/c17-15(18)10-3-1-9(2-4-10)13-7-11-5-6-12(16(19)20)8-14(11)21-13;;/h1-8,21H,(H3,17,18)(H3,19,20);2*1H

InChI key

FPNZBYLXNYPRLR-UHFFFAOYSA-N

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General description

DAPI (4′,6-Diamidino-2-phenylindole dihydrochloride) is a cell permeable, fluorescent dye that binds to DNA.
heat or sonication may be required. Solutions stored in the dark at room temperature or 4 °C should be stable for 2 to 3 weeks.

Application

DAPI is several times more sensitive than ethidium bromide for staining DNA in agarose gels. It may be used for photofootprinting of DNA, to detect annealed probes in blotting applications by specifically visualizing the double-stranded complex, and to study the changes in DNA and analyze DNA content during apoptosis using flow cytometry. DAPI staining has also been shown to be a sensitive and specific detection method for mycoplasma.
Suitable for
  • DNA staining in agarose gels
  • analysis of changes in DNA during apoptosis
  • detection of mycoplasma
  • photofootprinting of DNA
  • immunofluorescent staining of cells

DAPI has been used:-
  • in rapid monitoring of microbial contamination
  • in chromosomal banding technique
  • in detection of apoptotic cells
  • in fluorescence microscopy to track the DisA (DNA integritiy scanning protein) movement on Bacillus subtilis DNA
  • to stain mature pollen grains(0.5 mg/ml)

Biochem/physiol Actions

Cell permeable fluorescent minor groove-binding probe for DNA. Binds to the minor groove of double-stranded DNA (preferentially to AT rich DNA), forming a stable complex which fluoresces approximately 20 times greater than DAPI alone.

Caution

Protect from light.

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Description
Pricing

Pictograms

Exclamation mark

Signal Word

Warning

Hazard Statements

Hazard Classifications

Skin Irrit. 2 - Skin Sens. 1A - STOT SE 3

Target Organs

Respiratory system

WGK

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Personal Protective Equipment

dust mask type N95 (US), Eyeshields, Gloves

Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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  1. Which document(s) contains shelf-life or expiration date information for a given product?

    If available for a given product, the recommended re-test date or the expiration date can be found on the Certificate of Analysis.

  2. How do I get lot-specific information or a Certificate of Analysis?

    The lot specific COA document can be found by entering the lot number above under the "Documents" section.

  3. How should D9542, 4´,6-Diamidino-2-phenylindole dihydrochloride (DAPI), be dissolved?

    This product is soluble in water at 20 mg/mL, but may require heat.  DAPI does not dissolve readily in PBS, 0.1 M phosphate, or 0.1 M Tris (pH 7).

  4. How should D9542, 4´,6-Diamidino-2-phenylindole dihydrochloride (DAPI), be stored once in solution, and for how long would it be good?

    Solutions stored in the dark at room temperature have been stable for 2-3 weeks, but solutions exposed to light for the same time period have shown considerable degradation.  At 1 mg/mL, solutions are stable in the dark at 2-8 °C for several weeks.  For long-term storage, the solutions can be divided into aliquots and stored at -20 °C.

  5. Aside from the applications described on the product page, how is D9542, 4´,6-Diamidino-2-phenylindole dihydrochloride (DAPI), used?

    DAPI is used as a nuclear counterstain in our FISH protocol at 2 μg/mL in antifade mounting media. DAPI can also be used as a vital dye to stain mature pollen grains (0.5 μg/mL). DAPI can bind to RNA, but its selectivity for DNA over RNA is high.  It appears that DAPI uses an intercalating binding mode with RNA that is AU selective (Biochem., 31, 3103 (1992)). Selectivity of DAPI for DNA over RNA is greater than that shown by ethidium bromide and propidium iodide (Nucleic Acid Research, 6, 3535 (1979))

  6. How do I find price and availability?

    There are several ways to find pricing and availability for our products. Once you log onto our website, you will find the price and availability displayed on the product detail page. You can contact any of our Customer Sales and Service offices to receive a quote.  USA customers:  1-800-325-3010 or view local office numbers.

  7. What is the Department of Transportation shipping information for this product?

    Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product. 

  8. What is the stability of  D9542, 4´,6-Diamidino-2-phenylindole dihydrochloride?

    Product D9542, 4´,6-Diamidino-2-phenylindole dihydrochloride has a stability of 3 years as supplied. The product in solution is also stable in the dark at 4°C and showed no signs of degradation after three weeks at room temperature in the dark.  Material exposed to light showed significant degradation. We have developed an HPLC method for our DAPI products that shows reassay results from July 2012 for lots that were originally released for sale in December 2008, for both D9542 (DAPI dihydrochloride) and D9564 (DAPI dilactate).

  9. My question is not addressed here, how can I contact Technical Service for assistance?

    Ask a Scientist here.

Michal Bejerano-Sagie et al.
Cell, 125(4), 679-690 (2006-05-23)
In response to DNA damage, cells activate checkpoint signaling cascades to control cell-cycle progression and elicit DNA repair in order to maintain genomic integrity. The sensing and repair of lesions is critical for Bacillus subtilis cells entering the developmental process
Yan Li et al.
Toxicology letters, 154(3), 225-233 (2004-10-27)
Excessive exposure to synthetic and endogenous estrogens has been associated with the development of cancer in several tissues including the breast. 4-Hydroxyequilenin (4-OHEN), a major catechol metabolite of equine estrogens present in Premarin, an estrogen replacement formulation, has been shown
K Nakajima et al.
Letters in applied microbiology, 40(2), 128-132 (2005-01-13)
To apply fluorescent staining method for fast assessment of microbial quality of herbal medicines. The number of total bacteria and esterase-active bacteria on powdered traditional Chinese medicines were enumerated by fluorescent staining method using 6-carboxyfluorescein diacetate (6CFDA) and 4',6-diamidino-2-phenylindole (DAPI)
Yona Goldshmit et al.
Brain and behavior, 4(2), 187-200 (2014-04-01)
A major impediment for recovery after mammalian spinal cord injury (SCI) is the glial scar formed by proliferating reactive astrocytes. Finding factors that may reduce glial scarring, increase neuronal survival, and promote neurite outgrowth are of major importance for improving
J Kapuscinski
Biotechnic & histochemistry : official publication of the Biological Stain Commission, 70(5), 220-233 (1995-09-01)
DAPI (4',6-diamidino-2-phenylindole) is a DNA-specific probe which forms a fluorescent complex by attaching in the minor grove of A-T rich sequences of DNA. It also forms nonfluorescent intercalative complexes with double-stranded nucleic acids. The physicochemical properties of the dye and

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