10236276001
Roche
DAPI
4′,6-Diamidine-2′-phenylindole dihydrochloride
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
MDL number:
UNSPSC Code:
41116100
PubChem Substance ID:
Quality Level
Assay
>90%
form
powder
packaging
pkg of 10 mg
manufacturer/tradename
Roche
λmax
340 nm in aq. suspension
fluorescence
λex 340 nm; λem 488 nm (nur DAPI)
λex 364 nm; λem 454 nm (DAPI-DNA-Komplex)
storage temp.
room temp
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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Application
DAPI is a fluorescent dye that binds selectively to double-stranded DNA and forms strongly fluorescent DNA-DAPI complexes with high specificity. It is commonly used to detect mycoplasma in cell culture via fluorescence microscopy.
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.
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.
Quality
Purity: >90% (from N)
Principle
The fluorescent dye DAPI binds selectively to DNA and forms strongly fluorescent DNA-DAPI complexes with high specificity. DAPI, once added to tissue culture cells, is rapidly taken up into cellular DNA, yielding highly fluorescent nuclei and no detectable cytoplasmic fluorescence. When the cells are contaminated with Mycoplasmas, characteristic discrete fluorescent foci are readily detected over the cytoplasm and sometimes in intercellular spaces.
Preparation Note
Working solution: Solubility: 25 mg/ml in water
Preparation of stock solution
Dissolve in double-dist. water to a final concentration of 1 to 5 mg/ml.
Note: Do not use any buffers.
Preparation of working solution
Dilute the stock solution with methanol to a final concentration of 1 μg/ml. The working solution is stable at 2 to 8 °C for about 6 months.
Storage conditions (working solution): Stock solution (1 to 5 mg/ml) at -15 to -25 °C for 12 months.
Working solution (1μg/ml) at 2 to 8 °C for about 6 months.
Preparation of stock solution
Dissolve in double-dist. water to a final concentration of 1 to 5 mg/ml.
Note: Do not use any buffers.
Preparation of working solution
Dilute the stock solution with methanol to a final concentration of 1 μg/ml. The working solution is stable at 2 to 8 °C for about 6 months.
Storage conditions (working solution): Stock solution (1 to 5 mg/ml) at -15 to -25 °C for 12 months.
Working solution (1μg/ml) at 2 to 8 °C for about 6 months.
Reconstitution
In 2 to 10 ml double-dist. water; 1 to 5 mg/ml final concentration.
Note: Prepare aliquots and store at -15 to -25 °C.
Note: Prepare aliquots and store at -15 to -25 °C.
Storage Class Code
11 - Combustible Solids
WGK
WGK 2
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
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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