Ensuring that weighing instruments are calibrated is essential for accurate measurements. This approach should be acceptable for preparing the desired concentrations.
D9542
DAPI
for nucleic acid staining
Sinonimo/i:
4′,6-diamidino-2-fenilindolo, 2-(4-amidinofenil)-6-indolocarbammidina, DAPI
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Informazioni su questo articolo
Formula empirica (notazione di Hill):
C16H15N5 · 2HCl
Numero CAS:
Peso molecolare:
350.25
UNSPSC Code:
12352200
NACRES:
NA.52
PubChem Substance ID:
EC Number:
249-186-7
Beilstein/REAXYS Number:
4894417
MDL number:
Prezzi e disponibilità al momento non sono disponibili
Servizio Tecnico
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Permettici di aiutartigrade
Molecular Biology
Quality Segment
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
General description
DAPI (4′,6-diamidino-2-fenilindolo diidrocloruro) è un colorante fluorescente permeabile alle cellule che si lega al DNA.
Potrebbe essere necessario ricorrere a calore o trattamento con ultrasuoni. La soluzione conservata al buio a temperatura ambiente o 4 °C dovrebbe essere stabile per 2-3 settimane.
Application
Adatto per
DAPI trova impiego:-
- Colorazione del DNA in gel di agarosio
- Analisi di cambiamenti nel DNA durante l′apoptosi
- Rilevamento di micoplasma
- fotoimpronte del DNA
- Colorazione delle cellule per immunofluorescenza
DAPI trova impiego:-
- nel monitoraggio rapido di contaminazione microbica
- nella tecnica di rivelazione a bande dei cromosomi
- nella rivelazione di cellule apoptotiche
- Nella microscopia in fluorescenza per seguire il movimento del DisA (proteina per la scansione dell′integrità del DNA)) sul DNA di Bacillus subtilis
- Per colorare granuli di polline maturo (0,5 mg/mL)
DAPI è parecchie volte più sensibile del bromuro di etidio per colorare il DNA in gel di agarosio.
DAPI è parecchie volte più sensibile del bromuro di etidio per colorare il DNA in gel di agarosio. Può essere impiegato per fotoimpronte del DNA, rivelare sonde ibridizzate in applicazioni blotting visualizzando specificamente il complesso a doppio filamento, studiare i cambiamenti nel DNA e analizzare con la citometria di flusso il contenuto di DNA durante l′apoptosi. La colorazione via DAPI si è dimostrata un metodo di rivelazione sensibile e specifico dei micoplasma.
Biochem/physiol Actions
Sonda fluorescente per DNA permeabile alle cellule che si attacca al solco minore. Si attacca al solco minore del DNA a doppio filamento (preferenzialmente a DNA ricco di T) a formare un complesso stabile con una fluorescenza circa 20 volte maggiore del DAPI da solo.
Disclaimer
Tenere al riparo dalla luce.
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Questo articolo | |||
|---|---|---|---|
| description for nucleic acid staining | description 4′,6-Diamidine-2′-phenylindole dihydrochloride | description Cell-permeable DNA-binding dye. | description For Nuclear counterstain in immunoflourecence microscopy, High Content Screening (HCS), Chromosome staining and flow cytometry (FACS)., 1 mg/mL |
| grade Molecular Biology | grade - | grade - | grade - |
| technique(s) transfection: suitable | technique(s) - | technique(s) - | technique(s) transfection: suitable |
| suitability suitable for fluorescence | suitability - | suitability - | suitability suitable for nucleic acid staining |
| fluorescence λex 340 nm; λem 488 nm (nur DAPI), λex 364 nm; λem 454 nm (DAPI-DNA-Komplex) | fluorescence λex 340 nm; λem 488 nm (nur DAPI), λex 364 nm; λem 454 nm (DAPI-DNA-Komplex) | fluorescence λex ~359 nm, λem ~461 nm | fluorescence λex 364 nm; λem 454 nm (DAPI-DNA-Komplex), λex 340 nm; λem 488 nm (nur DAPI) |
| form powder | form powder | form solid | form liquid |
| solubility H2O: 20 mg/mL, PBS: insoluble | solubility - | solubility methanol: soluble, water: 5 mg/mL | solubility water: soluble |
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Parola di avvertenza
Warning
Codici di pericolo
Avvertenze di sicurezza
Hazard Classifications
Skin Irrit. 2 - Skin Sens. 1A - STOT SE 3
Organi bersaglio
Respiratory system
Classe di stoccaggio
11 - Combustible Solids
Che succede?
WGK 3
Punto di infiammabilità (°F)
Not applicable
Punto di infiammabilità (°C)
Not applicable
DPI (dispositivi di protezione individuale)
Eyeshields, Gloves, type N95 (US)
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Can I dissolve 1mg in 50ul of H2O to adjust the concentration to 20mg/ml, then dilute it to 0.5ug/ml~5ug/ml and apply it to the cells?
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Can this product be used to stain bacteria (E.coli) chromosome?
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DAPI is suitable for staining bacterial chromosomes, including those of E. coli.
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What is the working dilution of DAPI that should be used to stain the nuclei of human cells?
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Typical working concentration of DAPI used in most applications is 0.1μg/mL-10μg/mL. Please see the link below to review the product datasheet:
https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/774/984/d9564pis.pdfPlease see the link below to review a sample protocol from Cold Spring Harbor:
https://cshprotocols.cshlp.org/content/2011/1/pdb.prot5556.longUtile?
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what kind of the water used to dissolve DAPI? and what's the temperature should be dissolve the water?
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This product is soluble at 20 mg/mL in water at ambient temperature. No specific water type or treatment is required. Deionized water is typically used for laboratory applications.
Please see the link below to review the product data sheet:
https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/392/209/d9542pis.pdfUtile?
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What is the suggested concentration for product D9542-1MG?
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The suggested concentration for product D9542-1MG, DAPI, is in the range of 0.5 to 5 ug/ml.
Utile?
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Can the stain concentration be used for staining fish scales and any animal cells for ICC?
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Yes, DAPI can be used to stain fish scale nuclei. While specific data regarding the concentration and duration for staining fish scale nuclei is not available, it is suggested to start with a working concentration of around 0.5 mg/ml and optimize the staining for each specific cell type, as different cells may uptake the dye at varying rates. It is also advisable to try staining at both higher and lower concentrations. For staining mammalian cells, lower concentrations are typically used.
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What is the suitable DAPI for immunofluorescence staining of free-floating rat brain sections?
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Several DAPI products are suitable for staining free-floating rat brain sections. The same DAPI used for staining mycoplasma, cultured cells, or other cell types can be used for this purpose. Any of the DAPI products listed on the website should be suitable for this use.
Utile?
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肝細胞を染める染色プロトコルを教えてください。
1 risposta-
Please see the link below to review the product datasheet, which includes basic protocols:
https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/392/209/d9542pis.pdfA protocol specifically for hepatocyte staining is not available. However, the publication shown at the link below may be helpful:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5671579/Utile?
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Aside from the applications described on the product page, how is D9542, 4´,6-Diamidino-2-phenylindole dihydrochloride (DAPI), used?
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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))
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What is the stability of D9542, 4´,6-Diamidino-2-phenylindole dihydrochloride?
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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).
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