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PUREBLU™ DAPI

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PUREBLU™ DAPI thumbnail image 1 PUREBLU™ DAPI thumbnail image 2 PUREBLU™ DAPI thumbnail image 3 PUREBLU™ DAPI thumbnail image 4 PUREBLU™ DAPI thumbnail image 5
PUREBLU™ DAPI gallery image 1
HeLa cells were treated with BrdU for 3 hours and stained with mouse anti-BrdU antibody, clone Bu20a (MCA2483). Goat anti mouse IgG (H/L) DyLight® 549 conjugated antibody (red) (STAR117D549GA) was used as the secondary antibody. Cytoplasm was stained with rabbit anti-GAPDH antibody (AHP1628). As the secondary antibody, sheep anti rabbit IgG DyLight® 488 conjugated antibody (green) (STAR36D488GA) was used. PureBlu™ DAPI (1351303) was used as nuclear counterstain (blue)
PUREBLU™ DAPI gallery image 2
HeLa cells, formaldehyde fixed and permeabilized, stained with rat anti alpha tubulin (red) (MCA78G), mouse anti human CD49a, green (MCA1133) and PureBlu DAPI, blue (1351303).
PUREBLU™ DAPI gallery image 3
Paraffin section of human pancreas stained with guinea pig anti insulin (red) (5330-0104G), mouse anti chromogranin A, green (MCA4773) and PureBlu DAPI, blue (1351303).
PUREBLU™ DAPI gallery image 4
Paraffin section of human colon adenocarcinoma (HIER citrate pH6 antigen retrieval) blocked with 10% FCS and stained with mouse anti human cytokeratin 18, green (MCA1864H) and detected with goat anti mouse IgG:A488 (STAR117D488GA). The tissue was also counterstained with PureBlu DAPI, blue (1351303).
PUREBLU™ DAPI gallery image 5
HeLa cells were treated with 10 µg BrdU for 1 hour (B) or left untreated (A). Cells were stained with Mouse anti BrdU antibody, clone Bu20a (MCA2483) at a dilution of 1/25. As a secondary antibody, goat anti mouse IgG (H/L) DyLight® 549 conjugated antibody, red (STAR117D549GA) was used at a 1/50 dilution. Cytoplasm was stained with rabbit anti-GAPDH antibody (AHP1628) at a dilution of 1/100. As a secondary antibody, sheep anti rabbit IgG DyLight® 488 conjugated antibody, green (STAR36D488GA) was used at a 1/50 dilution. PureBlu™ DAPI, blue (1351303) was used as nuclear counterstain.

PUREBLU™ DAPI

Product Type
Accessory Reagent
Product Code Applications Pack Size List Price Quantity
1351303
Datasheet
F IF
MSDS
250 µg

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.

Product Details

Reconstitution
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

Storage Information

Storage
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.
Guarantee
Guaranteed until date of expiry. Please see product label.

More Information

Regulatory
For research purposes only

Applications of PUREBLU™ DAPI

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.
Application Name Verified Min Dilution Max Dilution
Flow Cytometry 1/500
Immunofluorescence 1/500
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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