DAPI is a cell permeable fluorescent compound (MW 350.3) that is able to stain the DNA of eukaryotic and prokaryotic cells by binding with high affinity to the minor groove of AT-rich DNA sequences. When DAPI is bound to DNA and excited by an ultraviolet light source, blue fluorescent emission can be detected with maximum emission at at 461 nm. PureBlu DAPI has a characteristic Stokes shift of approximately 100 nm, which makes this dye an optimal choice when good spectral separation is desired. PureBlu DAPI is compatible with fixed and unfixed cells.
- 1. Reconstitute one vial of lyophilized PureBlu DAPI Dye with 500 μl of de-ionized water, then vortex briefly to make 100x stock solution.
2. Dilute the 100x stock solution 1:100 with 1x phosphate buffered saline to make a 1 μg/ml staining solution.
- Reagents in the Kit
- 5 vials, 50 μg each, DAPI nuclear staining dye powder
- Prior to reconstitution store at - 20oC
After reconstitution store at +4oC or at -20oC if preferred
This product is photosensitive and should be protected from light. PureBlu™ DAPI Dye is stable for 12 months from date of reconstitution if stored at - 20oC or 6 months at 2-8oC.
- Guaranteed until date of expiry. Please see product label.
- For research purposes only
This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.
Applications of PUREBLU™ DAPI
Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.
- Instructions For Use
- Note: The optimal concentration for different cell types should be determined empirically.
1. Culture cells of interest under appropriate conditions.
2. Rinse cells with 1x phosphate buffered saline.
3. Optional: Fix cells with 3.7% formaldehyde at room temperature (18-25oC) for 10 minutes.
4. Optional: Rinse cells with 1x phosphate buffered saline and permeabilize with 0.1% Triton x-100 in phosphate buffered saline at room temperature (18-25oC) for 5 minutes.
5. Rinse cells with 1x phosphate buffered saline.
6. Stain with 1x staining solution at room temperature (18-25oC) for 15 minutes.
7. Rinse cells with 1x phosphate buffered saline.
8. Optional: Remove phosphate buffered saline and mount cells in antifade-mounting media.
9. Image cells.
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