Search Thermo Fisher Scientific
Search Thermo Fisher Scientific
A popular nuclear and chromosome counterstain, DAPI (4′,6-diamidino-2-phenylindole) emits blue fluorescence upon binding to AT regions of DNA.
1. Add 2 mL of deionized water (diH2O) or dimethylformamide (DMF) to the entire contents of the DAPI vial to make a 14.3 mM (5 mg/mL) DAPI stock solution. Note: DAPI has poor solubility in water, so sonicate as necessary to dissolve. The 5 mg/mL DAPI stock solution may be stored at 2–6°C for up to 6 months or at ≤–20°C for longer periods. |
2. Add 2.1 µL of the 14.3 mM DAPI stock solution to 100 µL PBS to make a 300 µM DAPI intermediate dilution |
3. Dilute the 300 µM DAPI intermediate dilution 1:1,000 in PBS as needed to make a 300 nM DAPI stain solution. |
First, fix and permeabilize cultured cells with a protocol appropriate for your sample.
1. Wash the cells 1–3 times in PBS as needed. |
2. Add sufficient 300 nM DAPI stain solution to cover the cells. |
3. Incubate for 1–5 minutes, protected from light. |
4. Remove the stain solution. |
5. Wash the cells 2–3 times in PBS. |
6. Image the cells. |
DAPI | |
---|---|
Excitation/Emission (nm) | 358/461 |
Standard filter set | DAPI |
EVOS® Light Cube | DAPI |
Storage conditions | ≤–20°C |
Drosophila melanogaster embryo staining. Wild type (Canton-S) Drosophila melanogaster embryo exhibiting microtubule staining (green fluorescence), denticle band staining (red fluorescence), and nuclear staining (blue-fluorescent DAPI).