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

DAPI (4',6-Diamidino-2-phenylindole) dihydrochloride (solution) 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).
Solvent and concentration: ddH2O: 5 mg/mL

Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs).
General Protocol
1. Preparation of DAPI working solution
1.1 Preparation of DAPI working solution
Dilute the stock solution in serum-free cell culture medium or PBS to obtain 1-10 ug/mL of working solution.
Note: Please adjust the concentration of DAPI working solution according to the actual situation.
2. Cell staining
2.1 Suspension cells（6-well plate）
a. 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 1×10⁶/mL.
b. Add 1 mL of working solution, and then incubate at room temperature for 3-10 minutes.
c. Centrifuge at 400 g at 4°C for 3-4 minutes and then discard the supernatant.
d. Wash twice with PBS, 5 minutes each time.
e. Resuspend cells with serum-free cell culture medium or PBS. Observation by fluorescence microscopy or flow cytometry.
2.2 Adherent cells
a. Culture adherent cells on sterile coverslips.
b. Remove the coverslip from the medium and aspirate excess medium.
c. Add 100 μL of working solution, gently shake it to completely cover the cells,and then incubate at room temperature for 3-10 minutes.
d. Wash twice with medium, 5 minutes each time.Observation by fluorescence microscopy or flow cytometry.

Storage
-20°C, 1 year. Protect from light

Precautions 1. Please adjust the concentration of DAPI working solution according to the actual situation.
2. This product is for R&D use only, not for drug, household, or other uses.
3. For your safety and health, please wear a lab coat and disposable gloves to operate.

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

350.25

C₁₆H₁₇Cl₂N₅

28718-90-3

Liquid

Light yellow to yellow

N=C(C1=CC2=C(C=C1)C=C(C3=CC=C(C(N)=N)C=C3)N2)N.Cl.Cl

Room temperature in continental US; may vary elsewhere.

Please store the product under the recommended conditions in the Certificate of Analysis.

• [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. Sep;70(5):220-33.  [Content Brief]