PUREBLU™ DAPI
PureBlu DAPI fluorescent DNA binding reagent (1351303) used to visualize nuclei of human circulating tumor cells by immunofluorescence in combination with cell surface markers.
Image caption:
Confocal microscopy representative images of isolated CTCs stained for epithelial and hematopoietic markers. Representative image by confocal microscopy of circulating cancer cells stained with anti-human EPCAM, anti-human pan-CK or anti-human CD45 Abs followed by goat anti-mouse secondary antibody Alexa 594-conjugated. DAPI was used to counteract nuclei. Magnification 100×. CK = cytokeratin, an epithelial cytoplasmic marker; DAPI = nuclear marker; EpCAM = epithelial membrane marker; CD45 = leukocyte common antigen.
From: Morelli MB, Amantini C, Rossi de Vermandois JA, Gubbiotti M, Giannantoni A, Mearini E, Maggi F, Nabissi M, Marinelli O, Santoni M, Cimadamore A, Montironi R, Santoni G.
Correlation between High PD-L1 and EMT/Invasive Genes Expression and Reduced Recurrence-Free Survival in Blood-Circulating Tumor Cells from Patients with Non-Muscle-Invasive Bladder Cancer.
Cancers (Basel). 2021 Nov 28;13(23):5989.
doi: 10.3390/cancers13235989.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
PureBlu DAPI fluorescent DNA stain (1351303) used to stain nuclei of RAW 264.7 cells by immunofluorescence.
Image caption:
Effect of astaxanthin on F-actin ring size. (A) Cells were treated similarly to the TRAP activity experiment, which was followed by staining with phalloidin to stain cytoplasm as well as DAPI to stain nuclei. F-actin rings are indicated by white arrows in the figures. (B) F-actin ring size per osteoclast cells. All data are shown as means ±SEM, ** p <0.01, Tukey–Kramer test. The bar in the figure represents 50μm.
From: Mamun-Or-Rashid, A.N.M.; Lucy, T.T.; Yagi, M.; Yonei, Y.
Inhibitory Effects of Astaxanthin on CML-HSA-Induced Inflammatory and RANKL-Induced Osteoclastogenic Gene Expression in RAW 264.7 Cells.
Biomedicines 2022, 10, 54.
doi: 10.3390/biomedicines10010054
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
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
- Reconstitute one vial of lyophilized PureBlu DAPI Dye with 500 μl of de-ionized water, then vortex briefly to make a 100x stock 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
- Acknowledgements
- Triton is a trademark of Dow Chemical Company
- Regulatory
- For research purposes only
Applications of PUREBLU™ DAPI
| 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
- Note: The optimal concentration for different cell types should be determined empirically.
Staining Procedure - Flow Cytometry Cell Viability
1. Grow cells of interest under conditions specific for the cell type. Harvest cells and obtain a single cell suspension.
2. Check cell numbers and viability using trypan blue.
3. Resuspend cells at 2 x 106 cells/ml in 0.5ml Staining Buffer (BUF073) or 1x PBS.
4. Add 5μl of 100x PureBlu DAPI Dye stock solution to 0.5ml cells.
5. Stain at room temperature for 15 min in the dark.
6. Optional: Rinse cells with Staining Buffer (BUF073) or 1x PBS.
7. Proceed with analysis by flow cytometry. When bound to double-stranded DNA (dsDNA) PureBlu DAPI Dye can be excited with either a 355 nm or a 405 nm laser, with optimum emission at 461nm.
Staining Procedure - Flow Cytometry Cell DNA Content
1. Grow cells of interest under conditions specific for the cell type. Harvest cells and obtain a single cell suspension.
2. Check cell numbers and viability using trypan blue.
3. Wash cells in Staining Buffer (BUF073) or 1x PBS.
4. Fix in ice cold 70% ethanol for 2 hr at 4°C.
5. Wash cells in Staining Buffer (BUF073) or 1x PBS.
6. Stain cells with 5μl of PureBlu DAPI Dye stock solution to 0.5 ml cells at a concentration of 2 x 106 cells/ml in cell staining buffer containing 0.1% Triton X-100.
Note: Optimal concentrations of cells and PureBlu DAPI may vary depending on cell type and should be determined through careful titration prior to final experimental analysis.
7. Stain at room temperature for 30 min in the dark.
8. Optional: Rinse cells with Staining Buffer (BUF073) or 1x PBS.
9. Proceed with analysis by flow cytometry. When bound to double-stranded DNA (dsDNA) PureBlu DAPI Dye can be excited with either a 355 nm or a 405 nm laser, with optimum emission at 461nm.
Staining Procedure - Cell Nuclear Visualization in Microscopy and Cell Imaging
1. Grow cells of interest under conditions specific for the cell type.
2. Rinse cells with 1x PBS.
3. Optional: Rinse cells with 1x PBS and permeabilize them with 1x PBST (0.1% Triton X-100 in 1x PBS) at room temperature for 5 min.
5. Rinse cells with 1x PBS.
6. Stain with 1x staining solution (diluted with PBS) at room temperature for 15 min.
7. Rinse cells with 1x PBS.
8. Optional: Remove PBS and mount cells in antifade mounting media.
9. Image cells.
Copyright © 2022 Bio-Rad Antibodies (formerly AbD Serotec)
Useful Reagents Available
| Description | Product Code | Applications | Pack Size | List Price | Quantity |
|---|---|---|---|---|---|
| Staining Buffer | BUF073 | F | 500 ml |
Product Specific References
References for PUREBLU™ DAPI
-
She, D.T. et al. (2018) SIRT2 Inhibition Confers Neuroprotection by Downregulation of FOXO3a and MAPK Signaling Pathways in Ischemic Stroke.
Mol Neurobiol. 55 (12): 9188-203. -
Carter-Timofte, M.E. et al. (2021) Antiviral Potential of the Antimicrobial Drug Atovaquone against SARS-CoV-2 and Emerging Variants of Concern.
ACS Infect Dis. acsinfecdis.1c00278. -
Earnest, K.G. et al. (2021) Development and characterization of a DNA aptamer for MLL-AF9 expressing acute myeloid leukemia cells using whole cell-SELEX.
Sci Rep. 11 (1): 19174. -
Paterno, R. et al. (2021) Hippocampal gamma and sharp-wave ripple oscillations are altered in a Cntnap2 mouse model of autism spectrum disorder.
Cell Rep. 37 (6): 109970. -
Roux-Biejat, P. et al. (2021) Acid Sphingomyelinase Controls Early Phases of Skeletal Muscle Regeneration by Shaping the Macrophage Phenotype.
Cells. 10 (11): 3028. -
Nogueira-Rodrigues, J. et al. (2021) Rewired glycosylation activity promotes scarless regeneration and functional recovery in spiny mice after complete spinal cord transection.
Dev Cell. Dec 27 [Epub ahead of print]. -
Morelli, M.B. et al. (2021) Correlation between High PD-L1 and EMT/Invasive Genes Expression and Reduced Recurrence-Free Survival in Blood-Circulating Tumor Cells from Patients with Non-Muscle-Invasive Bladder Cancer.
Cancers (Basel). 13 (23)Nov 28 [Epub ahead of print]. -
Mamun-Or-Rashid, A.N.M. et al. (2021) Inhibitory Effects of Astaxanthin on CML-HSA-Induced Inflammatory and RANKL-Induced Osteoclastogenic Gene Expression in RAW 264.7 Cells
Biomedicines. 10 (1): 54.
Fluorescent Spectraviewer
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