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DAPI dihydrochloride

Cat. No. M5106

All AbMole products are for research use only, cannot be used for human consumption.

DAPI dihydrochloride Structure
Synonym:

4',6-Diamidino-2-phenylindole dihydrochloride

Size Price Availability Quantity
5mg USD 45  USD45 In stock
10mg USD 60  USD60 In stock
25mg USD 118  USD118 In stock
50mg USD 215  USD215 In stock
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Quality Control & Documentation
Biological Activity

DAPI dihydrochloride is a blue fluorescent nucleic acid stain, preferentially binds dsDNA and associates with the minor groove AT clusters. Association of DAPI with dsDNA increases fluorescence approximately 20-fold, with an emission maximum of 460 nm. The increase in fluorescence is believed to be due to displacement of water molecules from the minor groove and DAPI. DAPI can also bind RNA and emits at a longer wavelength of 500 nm, with only a 20% increase in fluorescence.


Protocols for Counterstaining with DAPI:

Immunofluorescence Staining
Note: Cells may be fixed by method of choice, such as with 4% paraformaldehyde. DAPI staining is done after staining for other markers. Fixation and permeabilization of cells is not necessary to counterstain with DAPI.

1. Fix cells using method of choice.
2. Incubate the cells with phosphate buffered saline (PBS) for 15 minutes.
3. Dilute the DAPI solution (we recommend to 300 nM) with PBS. Remove excess PBS from the slide and cover with DAPI solution, making sure the cells are completely covered.
4. Incubate for up to 5 minutes. Rinse the slide several times to remove all free DAPI. It is recommended to use a mounting medium with antifade reagent to reduce fluorescence quenching. The slide is ready to view under the microscope with appropriate filters.


Nuclear Staining for Flow Cytometry
Note: Cells may be fixed with 4% paraformadehyde or absolute ethanol.

1. Fix cells with absolute ethanol or 4% paraformaldehyde.
2. Centrifuge, discard the supernatant, and add 5 ml of PBS, allowing cells to rehydrate for 15 minutes.
3. Dilute the DAPI to 3 µM (recommended) in staining buffer.
4. Centrifuge cell suspension from step 3 above, discard supernatant, and add 1 ml of DAPI solution, then incubate at room temperature for 15 minutes.
5. Analyze by flow cytometry using an instrument with appropriate lasers.

Product Citations
Chemical Information
Molecular Weight 350.25
Formula C16H17Cl2N5
CAS Number 28718-90-3
Solubility (25°C) Water 5 mg/mL (Need ultrasonic)
Storage -20°C, protect from light, sealed
References

[1] Qunxiong Wu, et al. Reprod Sci. LncRNA HOXA-AS2 Activates the Notch Pathway to Promote Cervical Cancer Cell Proliferation and Migration

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Keywords: DAPI dihydrochloride, 4',6-Diamidino-2-phenylindole dihydrochloride supplier, Fluorescent Dye, inhibitors, activators

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