PUREBLU™ Hoechst 33342

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PUREBLU™ Hoechst 33342

Product Type
Accessory Reagent
Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
1351304 F IF 280 µg
Hoechst 33342 is a cell-permeable fluorescent compound 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 Hoechst 33342 is bound to DNA and excited by an ultraviolet light source, blue fluoresecent emission can be detected with a maximum emission at 461 nm. PureBlu Hoescht 33342 has a characteristic Stokes shift of approximately 100 nm, which makes this dye an optimal choice when a good spectral separation is required. PureBlu Hoescht 33342 is compatible with fixed and unfixed cells. It exhibits high permeability for live cell membranes and is optimal for live cell DNA staining.

Product Details

Reconstitution
1. Add 500 μl of de-ionized water to one tube of lyophilized PureBlu Hoechst 33342 Dye, then vortex briefly to make the 100x stock solution (1.1 μg/ml [2 μM]).

2. Dilute the stock solution 1:100 with growth media (for live cells) or 1x phosphate buffered saline (for fixed cells) to make the 1x staining solution.
Molecular Weight
561.93
Reagents in the Kit
5 vials, 56 μg each, Hoechst 33342 nuclear staining dye powder

Storage Information

Storage
Prior to reconstitution store at -20oC
After reconstitution store at -20oC or +4oC
This product is photosensitive and should be protected from light. PureBlu™ Hoechst 33342 is stable for 12 months from date of reconstitution if stored at 20oC or 6 months at 4oC.
Shelf Life
Please see label for expiry date.

More Information

Regulatory
For research purposes only

Applications of PUREBLU™ Hoechst 33342

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 1/100
Immunofluorescence 1/100
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
Staining of Live Cells

1. Grow cells of interest under conditions specific for the cell type.

2. Replace growth media with 1x staining solution (diluted in fresh growth media) and incubate at 37oC for 15 minutes.

3. Rinse cells with 1x phosphate buffered saline (prewarmed to 37oC).

4. Aspirate phosphate buffered saline and add fresh growth media to cells (prewarmed to 37oC).

5. Image cells.


Staining of Fixed Cells

1. Grow cells of interest under conditions specific for the cell type.

2. Rinse cells with 1x phosphate buffered saline.

3. Fix cells with 4% formaldehyde at room temperature for 10 minutes.

4. Optional: Rinse cells with 1x phosphate buffered saline and permeabilize them with 0.1% Triton X-100 in 1x phosphate buffered saline at room temperature for 5 minutes.

5. Rinse cells with 1x phosphate buffered saline.

6. Stain with 1x staining solution (diluted with phosphate buffered saline) at room temperature 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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