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

DAPI (4',6-Diamidino-2-phenylindole) 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. In addition, because DAPI can pass through intact cell membranes, it can be used to stain 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^[2] (Ex/Em = 356/451 nm).

Guide (The following is our recommended protocol; this guideline is for reference only and should be modified according to your specific requirements).
1. Preparation of DAPI Working Solution
Dilute the stock solution with serum-free cell culture medium or PBS to prepare a working solution at a concentration of 1-10 μg/mL.
Note: Adjust the concentration of the DAPI working solution as needed.
2. Cell Staining
2.1 Suspension Cells (96-well Plate)
a. Centrifuge at 1000 g for 3-5 min at 4°C, then discard the supernatant. Wash twice with PBS, 5 min each time. The cell density is 1×10⁶/mL.
b. Add 1 mL of working solution and incubate at room temperature for 3-10 min.
c. Centrifuge at 400 g for 3-4 min at 4°C, then discard the supernatant.
d. Wash twice with PBS, 5 min each time.
e. Resuspend cells with serum-free cell culture medium or PBS. Observe by fluorescence microscopy or flow cytometry.
2.2 Adherent Cells
a. Culture adherent cells on sterile coverslips.
b. Remove the coverslips from the culture medium and aspirate excess medium.
c. Add 100 μL working solution, shake gently to fully cover the cells, and incubate at room temperature for 3-10 min.
d. Wash twice with culture medium, 5 min each time. Observe under a fluorescence microscope or by flow cytometry.
Precautions 1. Adjust the concentration of DAPI working solution according to actual experimental conditions.
2. This product is for R&D use only, not for pharmaceutical, household or other applications.
3. For personal safety and health, please wear a lab coat and disposable gloves during operation.

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

277.32

C₁₆H₁₅N₅

47165-04-8

Solid

Light yellow to yellow

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

Room temperature in continental US; may vary elsewhere.

-20°C, protect from light

*In solvent : -80°C, 6 months; -20°C, 1 month (protect from 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 (protect from 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, et al. DAPI: a DNA-specific fluorescent probe. Biotech Histochem. 1995 Sep;70(5):220-33.  [Content Brief]

  [2]. Kuduk SD, et al. Amidine derived inhibitors of acid-sensing ion channel-3 (ASIC3). Bioorg Med Chem Lett. 2009 Aug 1;19(15):4059-63.  [Content Brief]