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CYTO-ID® Green long-term cell tracer kit

Live cell fluorescent labeling over extended time periods with no apparent toxic effects
 
ENZ-51036-K025 1 Kit 204.00 USD
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  • Allows dual labeling with a variety of CELLESTIAL® fluorescent probes
  • Minimal transfer of fluorescence from dye-labeled to unlabeled cells
  • Suitable for long-term cell viability, cytotoxicity, cell adhesion, cell migration and cell-cell fusion assays
CYTO-ID® Green long-term cell tracer kit uses proprietary noncovalent cell labeling technology to stably incorporate a green fluorescent dye containing hydrophobic aliphatic chains into the cell membrane’s lipid bilayer. The dye may be loaded into cells by following the included protocol. The labeling buffer is isotonic for mammalian cells and contains no detergents or organic solvents. The appearance of labeled cells may vary depending upon the cell type from uniformly bright to punctuate. This difference is thought to relate to the extent of membrane internalization occurring after cell labeling. The CYTO-ID® Green tracer dye fluorescence is independent of pH within normally encountered physiologic ranges and fluorescence intensity per cell is typically unaffected by the ultimate pattern of dye distribution. The CYTO-ID® Green tracer dye is not toxic to cells, as determined using the benchmark MTT cell viability assay. The dye is well retained by cells for up to 96 hours after loading, and is passed to daughter cells upon mitosis. Since the dye does not covalently modify proteins within the cells, normal physiological responses are better preserved than with molecular probes based upon thiol-reactive chloromethyl-based or amine-reactive succinimidyl ester-based fluorescent dyes. Dual labeling is also possible using a variety of available CELLESTIAL® dyes. Labeled cells can be visualized by epifluorescence or confocal fluorescence microscopy. Additionally, dye-labeled and unlabeled cell populations can be analyzed by flow cytometry. No transfer of fluorescence to adjacent cells was observed after a prolonged 96 hour incubation period. This is in stark contrast to Calcein AM and BCECF AM, which are only retained within viable cells for a few hours at physiological temperatures. The kit is suitable for a variety of applications including long term cell viability, cytotoxicity, cell adhesion, cell migration and cell-cell fusion studies.
ENZ-51036 Spectra
Figure 1. Fluorescence excitation (359, 460) and emission (527) spectra for the CYTO-ID® Green tracer dye. All spectra were determined for cell-bound dye.
ENZ-51036 image
Figure 2: Composite bright-field (A) and fluorescence microscopy images (B) demonstrating staining of Jurkat cells with CYTO-ID® Green Tracer dye. Standard FITC (Green) filter set was used to image the membrane-bound signal.
129L4-NS web
Figure 3: Flow cytometry analysis of fluorescence of mixed population of Jurkat cells over time. Jurkat cells stained with CYTO-ID® Green Tracer dye were mixed with an unstained population of Jurkat cells and incubated over a 120-hour period.
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ENZ-51036 Spectra ENZ-51036 image 129L4-NS web

Product Specification

Applications:Flow Cytometry
 
Application Notes:CYTO-ID® Green long-term cell tracer kit is suitable for a variety of applications including long term cell viability, cytotoxicity, cell adhesion, cell migration and cell-cell fusion studies.
 
Quality Control:A sample from each lot of CYTO-ID® Green long-term cell tracer kit is used to stain Jurkat cells and analyzed by flow cytometry, using the procedures described in the user manual.  Mean fluorescence of stained to unstained cells is greater than 5. 
 
Quantity:25 assays
 
Handling:Protect from light. Avoid freeze/thaw cycles.
 
Shipping:Shipped on Dry Ice
 
Short Term Storage:-20°C
 
Long Term Storage:-20°C
 
Kit/Set Contains:CYTO-ID® Green tracer dye, 50µl
4X Labeling Buffer, 12.5ml
10X HBSS, 25ml
 

General Literature References

Cell tracking 2007: A proliferation of probes and applications: P.K. Wallace & K.A. Muirhead; Immunol. Invest. 36, 527 (2007), Abstract;
In Living Color: Flow Cytometry and Cell Sorting Protocols: Poon, R.Y., et al; R.A Diamond & S. DeMaggio (eds.), Springer Verlag (New York), 302 (2000), Book,
Fluorescent cell labeling for in vivo and in vitro cell tracking: P.K. Horan, et al.; Methods Cell Biol. 33, 469 (1990), Abstract;
Stable Cell Membrane Labeling: P.K. Horan & S.E. Slezak; Nature 340, 167 (1989), Abstract;

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