Cell Explorer™ Live Cell Labeling Kit *Green Fluorescence with 405 nm Excitation*

Additional fluorescent color(s): 
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<p>Image of CPA cells in 96-well Costar black wall/clear bottom plate stained with Cell Explorer™ Live Cell Labeling Kit *Green Fluorescence with 405 nm Excitation*.</p>
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Unit Size: Cat No: Price (USD): Qty:
200 Tests 22615 $195


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Additional Ordering Information
Telephone: 1-800-990-8053
Fax: 1-408-733-1304
Email: sales@aatbio.com
International: See distributors





Overview

Ex/Em (nm)410/500
Storage Freeze (<-15 °C)
Minimize light exposure
InstrumentsFluorescence microplate reader, Flow cytometer
Category Cell Biology
Labeling Cells
Related Fluorescence Imaging
Our Cell Explorer™ fluorescence imaging kits are a set of tools for labeling cells for fluorescence microscopic investigations of cellular functions. The effective labeling of cells provides a powerful method for studying cellular events in a spatial and temporal context. This particular kit is designed to uniformly label live cells for the flow cytometric analysis of live cells with the violet laser (405 nm excitation). The kit uses a proprietary dye that gets enhanced fluorescence upon entering into live cells. The dye is a hydrophobic compound that easily permeates intact live cells. The hydrolysis of the weakly fluorescent substrate by intracellular esterases generates a strongly fluorescent hydrophilic product that is well-retained in the cell cytoplasm. It can be readily adapted for flow cytometry applications. The fluorescent dye used in the kit is well excited with the violet laser (405 nm excitation) to fluorescence at ~510 nm. The kit provides all the essential components with an optimized cell-labeling protocol. It is useful for a variety of studies, including cell adhesion, chemotaxis, multidrug resistance, cell viability, apoptosis and cytotoxicity.




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Protocol


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This protocol only provides a guideline, and should be modified according to your specific needs.
At a glance

Protocol summary

  1. Prepare cells in growth medium
  2. Remove growth medium
  3. Add CytoCalcein™ Violet 500 working solution (100 µL/well for a 96-well plate or 25 µL/well for a 384-well plate)
  4. Stain the cells at 37°C for 30 minutes to 1 hour
  5. Wash the cells
  6. Examine the specimen under fluorescence microscope or flow cytometer with a filter set at Ex/Em = 405/510 nm

Important notes
Thaw all the components at room temperature before opening.

Key parameters
Instrument:Fluorescence microplate reader
Excitation:405 nm
Emission:510 nm
Recommended plate:Solid black
  
Instrument:Flow cytometer
Excitation:405 nm
Emission:510 nm
Preparation of stock solution
Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles.

1. CytoCalcein™ Violet 500 stock solution:
Add 20 µL of DMSO into the vial of CytoCalcein™ Violet 500 (Component A) and mix well to make CytoCalcein™ Violet 500 stock solution. Protect from light. Note: 20 µL of CytoCalcein™ Violet 500 stock solution is enough for 1 plate.

Preparation of working solution

Add 20 µL of CytoCalcein™ Violet 500 stock solution into 10 mL of HHBS (Component B) and mix well to make CytoCalcein™ Violet 500 working solution.

For guidelines on cell sample preparation, please visit
https://www.aatbio.com/resources/guides/cell-sample-preparation.html

Sample experimental protocol
  1. Remove the growth medium.

  2. Add 100 µL/well (96-well plate) or 25 µL/well (384-well plate) CytoCalcein™ Violet 500 working solution into the cell plate.

  3. Incubate the cells in a 37°C, 5% CO2 incubator for 30 minutes to 1 hour.

  4. Remove the CytoCalcein™ Violet 500 working solution from the cells, wash the cells with HHBS (Component B) for 2 to 3 times, and replace with HHBS.

  5. Analyze the cells using a fluorescence microscope or flow cytometer with a filter set at Ex/Em = 405/510 nm. Note: Alternatively, cells can be fixed at this point for later imaging (fluorescence intensity might be decreased upon fixation).
Example data analysis and figures

Figure 1.

Image of CPA cells in 96-well Costar black wall/clear bottom plate stained with Cell Explorer™ Live Cell Labeling Kit *Green Fluorescence with 405 nm Excitation*.

Disclaimer
AAT Bioquest provides high-quality reagents and materials for research use only. For proper handling of potentially hazardous chemicals, please consult the Safety Data Sheet (SDS) provided for the product. Chemical analysis and/or reverse engineering of any kit or its components is strictly prohibited without written permission from AAT Bioquest. Please call 408-733-1055 or email info@aatbio.com if you have any questions.





References & Citations

Autophagy proteins are not universally required for phagosome maturation
Authors: Marija Cemma, Sergio Grinstein, John H Brumell
Journal: Autophagy (2016): 1440--1446

Differential detection of tumor cells using a combination of cell rolling, multivalent binding, and multiple antibodies
Authors: Ja Hye Myung, Khyati A Gajjar, Jihua Chen, Robert E Molokie, Seungpyo Hong
Journal: Analytical chemistry (2014): 6088--6094

Versatile fabrication of nanoscale sol--gel bioactive glass particles for efficient bone tissue regeneration
Authors: Bo Lei, Xiaofeng Chen, Xue Han, Jiaan Zhou
Journal: Journal of Materials Chemistry (2012): 16906--16913

Advanced glycation end-products increase IL-6 and ICAM-1 expression via RAGE, MAPK and NF-$\kappa$B pathways in human gingival fibroblasts
Authors: K Nonaka, Y Kajiura, M Bando, E Sakamoto, Y Inagaki, JH Lew, K Naruishi, T Ikuta, K Yoshida, T Kobayashi
Journal: Journal of Periodontal Research






Additional Documents

 
Safety Data Sheet (SDS)


Certificate of Analysis