PUREBLU™ Hoechst 33342
PureBlu Hoechst 33342 stain (1351304) used to lable nuclei of apoptotic HepG2 cells using immunofluorescence.
Image caption:
Apoptotic response of HepG2 cells after treatment with quercetin (Q), fingolimod (F) or combination of both drugs (Q + F). Apoptotic (white arrows in the insets) and non-apoptotic (gray arrows in the insets) nuclei were scored, and the percentage of apoptotic cells in each sample was determined. Bar, 100 µm. * p < 0.001.
From: Momchilova A, Nikolaev G, Pankov S, Vassileva E, Krastev N, Robev B, Krastev D, Pinkas A, Pankov R.
Effect of Quercetin and Fingolimod, Alone or in Combination, on the Sphingolipid Metabolism in HepG2 Cells.
Int J Mol Sci. 2022 Nov 11;23(22):13916.
doi: 10.3390/ijms232213916.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
PUREBLU™ Hoechst 33342 reagent used to label nuclei of HEK293T cells by immunofluorescence.
Image caption:
HA-0N3RAD HEK293 cells propagate AD-brain derived tau seeds. A) Representative immunofluorescent images of HA-0N3RAD cells at passage 4 (P4), probing with phosphorylation-specific antibodies: AT180, AT8, AT100, pS396, and pS422, and with an amyloid specific dye, Amytracker 680. sb = 10μm
From: Huang, M. & McEwan, W.A.
Sensitive detection and propagation of brain-derived tau assemblies in HEK293 based wild-type tau seeding assays.
bioRχiv (2024) 18 Jul [Preprint ahead of review].
doi: 10.1101/2024.07.18.604077.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
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P IF ResetPUREBLU™ Hoechst 33342
- Product Type
- Accessory Reagent
- Specificity
- PUREBLU™ Hoechst 33342
Antibody Quality and Performance Guarantee
| Hoechst 33342 is a cell-permeable fluorescent compound (MW 561.93) 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 fluorescent 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. |
- 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. - Reagents In The Kit
- 5 vials, 56 μg each, Hoechst 33342 nuclear staining dye powder
- Regulatory
- For research purposes only
- Guarantee
- Guaranteed until date of expiry. Please see product label.
Prior to reconstitution store at -20°C
After reconstitution store at -20°C or +4°C
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 -20°C or 6 months at +4°C.
After reconstitution store at -20°C or +4°C
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 -20°C or 6 months at +4°C.
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
-
1351304
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.
Triton is trademark of Dow Chemical Company
References for PUREBLU™ Hoechst 33342
-
Momchilova, A. et al. (2022) Effect of Quercetin and Fingolimod, Alone or in Combination, on the Sphingolipid Metabolism in HepG2 Cells.
Int J Mol Sci. 23 (22): 13916. -
Labrador-Garrido, A. et al. (2023) Live cell in situ lysosomal GCase activity correlates to alpha-synuclein levels in human differentiated neurons with LRRK2 and GBA1 mutations.
Front Cell Neurosci. 17: 1229213. -
Huang, M. & McEwan, A.W. (2024) Sensitive detection and propagation of brain-derived tau assemblies in HEK293 based wild-type tau seeding assays
bioRχiv: 18 Jul. [Epub ahead of print].
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