DAPI dilactate  (Synonyms: 4',6-Diamidino-2-phenylindole dilactate)

DAPI (4',6-Diamidino-2-phenylindole) dilactate is a DAPI dye. DAPI is a fluorescent dye that binds strongly to DNA. It binds to the AT base pair of the double-stranded DNA minor groove, and one DAPI molecule can occupy three base pair positions. The fluorescence intensity of DAPI molecules bound to double-stranded DNA is increased by about 20 times, and it is commonly observed with fluorescence microscopy, and the amount of DNA can be determined based on the intensity of fluorescence. DAPI cannot penetrate intact cell membranes and is commonly used for staining both live and fixed cells^[1]. DAPI (Compound 3) is an acid-sensing ion channel 3 (ASIC3) inhibitor. DAPI binds to ASIC3 and blocks the channel function. DAPI can be used in the study of chronic pain treatment (Ex/Em = 356/451 nm)^[2].

Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs).
1. Preparation of DAPI dilactate working solution
1.1 Preparation of stock solution
Dissolve DAPI dilactate in ddH₂O to obtain 5 mg /mL stock solution (DAPI, dilactate 10.9 mM).
Note: It is recommended to store the stock solution at -20°C or -80°C in the dark and avoid repeated freezing and thawing.
1.2 Preparation of DAPI dilactate working solution
The stock solution was diluted 1 : 5000 in PBS to 1 μg/mL. Store the working solution at 4 °C.
Note: Please adjust the concentration of DAPI dilactate working solution as needed
2. Cell staining (Suspension cells)
2.1 Centrifuge at 1000 g at 4°C for 3-5 minutes and then discard the supernatant. Wash twice with PBS, 5 minutes each time. The cell density is 1×10⁶mL.
2.2 Add 1 mL of working solution, and then incubate at room temperature for 3-10 minutes.
2.3 Centrifuge at 400 g at 4°C for 3-4 minutes and then discard the supernatant.
2.4 Wash twice with PBS, 5 minutes each time.
2.5 Resuspend cells with serum-free cell culture medium or PBS. Observation by fluorescence microscopy or flow cytometry.
3. Cell staining (Adherent cells)
3.1 Culture adherent cells on sterile coverslips.
3.2 Remove the coverslip from the medium and aspirate excess medium.
3.3 Add 100 μL of working solution, gently shake it to completely cover the cells, and then incubate at room temperature for 3-10 minutes.
3.4 Wash twice with medium, 5 minutes each time. Observation by fluorescence microscopy or flow cytometry.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

457.48

C₂₂H₂₇N₅O₆

28718-91-4

Solid

Light yellow to yellow

O=C(C(C)O)O.N=C(C1=CC=C(C2=CC3=CC=C(C(N)=N)C=C3N2)C=C1)N.O=C(C(C)O)O

Room temperature in continental US; may vary elsewhere.

-20°C, sealed storage, away from moisture and light

*In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light)

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

• [1]. Kapuscinski J. DAPI: a DNA-specific fluorescent probe. Biotech Histochem. 1995 Sep;70(5):220-33.  [Content Brief]

  [2]. Nazaroff CD, et al. Assessment of Lung Eosinophils In Situ Using Immunohistological Staining. Methods Mol Biol. 2021;2223:237-266.  [Content Brief]