DAPI (4′,6-Diamidino-2-Phenylindole, dihydrochloride) | 28718-90-3 Biotium貨號40043 (10mg/ml, 1ml)
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DAPI (4′,6-Diamidino-2-Phenylindole, dihydrochloride) | 28718-90-3 Biotium貨號40043 (10mg/ml, 1ml)
DAPI (4′,6-Diamidino-2-Phenylindole, dihydrochloride) 是一種流行的藍色 DNA 染料，用於固定細胞的經典藍色核複染劑(nuclear counterstain)。如螢光顯微鏡( fluorescence microscopy)、染色體染色(chromosome staining)和流式細胞術( flow cytometry)。該染料與 dsDNA 的小溝結合，螢光增強約 20 倍，對富含 A-T 的區域具有更高的親和力。以粉末或溶液形式提供，可用於更高濃度的活細胞染色。λEx/λEm（含 DNA）= 358/461 nm。儲存在 4°C 並避光。
DAPI的基本機制
Find the Right Nuclear Stain for Your Application
DAPI 可用於染色哺乳動物細(mammalian cells)以及革蘭氏陽性(gram-positive)和革蘭氏陰性細菌(gram-negative bacteria)。在酵母( yeast)中，染色很弱，沒有細胞核。 DAPI 的膜滲透性低於 Hoechst，通常用於固定細胞染色。Hoechst 染料具有膜滲透性，更常用於活細胞或固定細胞染色和細胞週期分析。雖然 Hoechst 和 DAPI 顯示出比嵌入 DNA 染料更低的細胞毒性，但它們在活細胞中結合 DNA 並具有潛在危險。 DAPI 和 Hoechst 通過 UV 激發進行光轉換以形成綠色螢光染料，這會導致多色成像中的偽影。有關詳細信息，請參閱我們的技術提示避免 DAPI 和 Hoechst 的 UV 光轉換產生的偽影。
DAPI 不能滲透活細胞，但很有用作為固定細胞或組織切片的核複染劑。在更高的濃度(~10 ug/mL)，DAPI 可用於活細胞染色。
| Product | Catalog Number | MW | Unit Size | Format |
|---|---|---|---|---|
| DAPI Powder (dilactate) |
40009 | 457.49 | 10 mg | Yellow solid |
| DAPI Powder (dihydrochloride) |
40011 | 350.25 | 10 mg | Yellow solid |
| DAPI Solution (dilactate) |
40043 | 457.49 | 1 mL (10 mg/mL in water) |
Yellow solution |
| CAS number | 28718-90-3 |
|---|---|
| Probe cellular localization | Nucleus |
| For live or fixed cells | For fixed cells, For live/intact cells |
| Detection method/readout | Fluorescence microscopy, Flow cytometry |
| Assay type/options | DNA content/cell cycle profiling by flow cytometry, Long term staining (24-72h), No-wash staining, Real-time imaging, Tissue staining |
| Cell permeability | Membrane permeant |
| Apoptosis/viability marker | All cell stain |
| Colors | Blue |
| Excitation/Emission | 358/461 nm (with DNA) |
| Storage Conditions | Store at 2 to 8 °C, Protect from light |
combined with other fluorescent probes.
2. Remove medium from the cells and replace with medium containing dye.
3. Incubate cells at room temperature or 37°C for 5-15 minutes, then image.
Note: Washing is not necessary for specific staining, but nuclear staining is
stable after washing.
1. Prepare 10X dye solution by diluting DAPI to 100 ug/mL in fresh, complete
culture medium. DAPI can be combined with other fluorescent probes, which
should be diluted to 10 times the final desired concentration.
2. Without removing the medium from the cells, add 1/10 volume of 10X dye
directly to the well.
3. Immediate mix thoroughly by gently pipetting the medium up and down.
For larger well sizes (e.g., 24-well to 6-well plates), the plate can be gently
swirled to mix.
4. Incubate cells at room temperature or 37°C for 5-15 minutes, then image.
Note: Washing is not necessary for specific staining, but nuclear staining is
stable after washing.
antibodies or other probes, and can be diluted in buffers with detergent or
blocking agents if convenient.
2. Add the PBS with dye to cells or tissue sections and incubate at room
temperature for at least 5 minutes.
3. Image the samples; washing is optional but not required.
Note: Samples can be stored at 4°C after staining and before imaging.
Note: DAPI can be included directly in antifade mounting medium for onestep mounting and staining. When using DAPI in mounting medium, longer
incubation times may be required for DAPI to completely penetrate the cell
nuclei.
be stained with 10 ug/mL DAPI in PBS or 150 mM NaCl for 30 minutes at room
temperature. DAPI tends to stain dead cells more brightly than live cells.
In S. cerevisiae, DAPI preferentially stains dead yeast with nuclear and
cytoplasmic staining when used at 10 ug/mL in PBS; in live yeast DAPI shows dim
mitochondrial staining.
Biotium 提供範圍廣泛的螢光探針用於研究凋亡細胞(Apotosis)
The NucView® Caspase-3 Substrate comes in 3 colors, and measures caspase activity in real time, in live cells. MitoView™ 633 can measure the decrease in mitochondria membrane potential that occurs in early apoptosis. CF® Dye Annexin V conjugates come in many color options, and label PS on the membranes of late apoptotic cells. We also offer kits containing combinations of these probes.