product description
DAPI is a blue fluorescent dye commonly used for nuclear staining, which preferentially binds to double stranded DNA, especially to the AT region in small grooves. In addition, DAPI can also bind to RNA, but the binding mode is different. Compared with DNA complexes, the fluorescence emission wavelength of DAPI/RNA complexes is longer (about 500 nm), while the emission wavelength of DAPI/DNA complexes is about 460 nm.
In multi-color fluorescence staining technology, DAPI is commonly used as a nuclear staining agent because its blue fluorescence can form a sharp contrast with green, yellow, or red fluorescence. DAPI has high nuclear specificity and almost no staining of cytoplasm. This dye solution has a concentration of 3 μ M and is not a sterile preparation.
ordering information
product name | Item number | specifications |
DAPI dye solution | AS21L122 | 10mL |
DAPI dye solution | AS21L122 | 50mL |
Transportation and storage
Blue ice transportation- Store at 20 ℃ away from light, with a shelf life of 12 months.
Usage
1. Re staining of adherent cells
1.1 Sample Preparation
1.1.1 Use appropriate fixatives to fix the adherent cell sample, ensuring that the cell morphology remains intact. After fixation, use DAPI dye for nuclear staining.
1.2 Re dyeing process
1.2.1 Temporarily equilibrate the sample with PBS;
1.2.2 Add an appropriate amount of DAPI staining solution to ensure complete coverage of the cells and incubate for 3-5 minutes;
1.2.3 Rinse the sample several times with PBS and gently remove excess liquid to avoid cell drying;
1.2.4 Observe the sample under a fluorescence microscope equipped with appropriate filters.
2. Re staining of suspended cells
2.1 Sample Preparation
2.2.1 Collect 2 × 105~1 × 106 suspended cells, collect the precipitate by centrifugation, and discard the supernatant;
2.2.2 Gently tap the test tube to resuspend the precipitate in the remaining liquid, and then add 1mL of PBS;
2.2.3 Slowly add the cell suspension to 4mL ethanol while vortexing at maximum speed. Place ethanol containing cells at -20 ° C for 5-15 minutes;
2.2.4 Centrifuge the cells to precipitate and discard the supernatant;
2.2.5 Gently tap the test tube to loosen the precipitate, then add 5ml of PBS.
2.2 Re dyeing process
2.2.1 Centrifuge the cells to precipitate, discard the supernatant, gently tap the test tube to loosen the precipitate, and add 2-3mL of DAPI staining solution;
2.2.2 Incubate at room temperature for 15 minutes;
2.2.3 If observing cells using a fluorescence microscope, centrifuge the sample, discard the supernatant, and resuspend the precipitate in fresh buffer. Add a drop of cell suspension onto the microscope slide, cover the slide and observe.
matters needing attention
1. This product is only for scientific experimental research and should not be used in clinical diagnosis, treatment, or other fields.
2. For your safety and health, please wear lab clothes and disposable gloves when operating.
3. Use a fluorescence microscope or laser confocal microscope to observe blue fluorescence.
4. Bring your own PBS, fixative, and anti fluorescence quenching sealing solution.
5. It is recommended to take 1-2 samples for preliminary experiments when using this kit for the first time.
Related product recommendations
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1X PBS (sterile) (item number: AC08L011)
This product is only for scientific experimental research and should not be used in clinical diagnosis, treatment, or other fields.
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