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Nuclear-ID® Blue/Red cell viability reagent (GFP-Certified®)

Distinguishes between live vs dead cells in a single reagent.
 
ENZ-53005-C100 100 µl 221.00 USD
 
The Nuclear-ID® Blue/Red cell viability reagent (GFP-Certified®) (Prod. No. ENZ-53005) is a mixture of a blue fluorescent cell-permeable nucleic acid dye and a red fluorescent cell-impermeable nucleic acid dye that is suited for staining dead nuclei. The staining pattern arising from the simultaneous combination of these two dyes permits determination of live and dead cell populations by fluorescence/confocal microscopy. The reagent, supplied as a 1000x solution, is sufficient for 1000 microscopy assays. The single-tube format makes this cell viability reagent easy to use. It leaves the cytoplasm unstained, potentially allowing visualization of other cell markers. This reagent is specificially designed for use with GFP-expressing cell lines.

Product Specification

Quantity:100μl (for 1000 microscopy assays).
 
Purity:≥93% (HPLC)
 
Short Term Storage:-20°C
 
Long Term Storage:-80°C
 
Handling:Protect from light. Avoid freeze/thaw cycles.
 
Miscellaneous/General:Application Note: Multiplexed Assay for IL-6 Secretion and Cell Viability Using an Epithelial Ovarian Cancer Cell Line

Application Note: Validation of a Novel Tumoroid-Based Cell Culture Model to Perform 3D in vitro Cell Signaling Analyses
 
Background / Technical Information:Wavelength Maxima
Live (Blue): Excitation: 350nm; Emission: 461nm
Dead (Red): Excitation: 571, 619nm; Emission: 639nm

The Nuclear-ID® Blue/Green cell viability reagent is a member of the CELLestial® product line, reagents and assay kits comprising fluorescent molecular probes that have been extensively benchmarked for live cell analysis applications. CELLestial® reagents and kits are optimal for use in demanding imaging applications, such as confocal microscopy, flow cytometry and HCS, where consistency and reproducibility are required.
 
Protocol:Wide Field Fluorescence/Confocal Microscopy
Reagent Preparation:
Mix 1μL of Nuclear-ID®  Blue/Red Cell Viability Reagent in 1 mL of buffer of choice. This volume is sufficient for 10 assays and may be scaled according to need.

Staining Adherent cells:
1. Grow cells directly onto glass slides or polystyrene tissue culture plates until ~80% confluent via standard tissue culture practices.
2. Remove growth media.
3. Dispense the freshly diluted staining solution in a volume sufficient for covering the cell monolayer.
4. Protect samples from light and incubate for 30 minutes at 37°C.
5. Remove the excess staining solution and, if necessary, add a few drops of buffer to prevent the cells from drying out.
6. Cover cells with a glass cover slip and observe under a fluorescence/confocal microscope with a dual filter set for DAPI (Ex/Em: 350/470nm) and Texas Red (Ex/Em: 540/605 nm). 

Staining Non-Adherent Cells:
1. Grow cells via standard tissue culture practices. 
2. Collect about 1 x 105 cells.  Centrifuge at 500 x g for 5 minutes.  Remove supernatant.
3. Re-suspend cells in a volume of the freshly diluted staining solution sufficient for covering the cell pellet.
4. Protect samples from light and incubate for 30 minutes at 37°C.
5. Centrifuge at 500 x g for 5 minutes. Remove supernatant.
6. Re-suspend cells in 100µL buffer.
7. Plate 10-15µL of cells on a glass slide.
8. Cover cells with a glass cover slip and observe under a fluorescence/confocal microscope with a dual filter set for DAPI (Ex/Em: 350/470nm) and Texas Red (Ex/Em: 540/605 nm). 

 
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Figure 1: Nuclear-ID® Blue/Red dye is detected as blue-stained nuclei in live cells and fluorescent-red nuclei in dead cells (inset, arrow).
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Product Literature References

Phenotypic switch in blood: effects of pro-inflammatory cytokines on breast cancer cell aggregation and adhesion: Y. Geng, et al.; PLoS One 8, e54959 (2013), Application(s): Breast cancer cell viability observed by confocal microscopy, Abstract; Full Text

General Literature References

Constitutive expression of the machinery for programmed cell death: M. Weil, et al.; J. Cell Biol. 133, 1053 (1996), Abstract;
Principles and Methods of Toxicology, Third Edition: A.W. Hayes, Ed.; Raven Press 1231 (1994), Book,

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