Cell Meter™ Cell Viability Assay Kit *NIR Fluorescence Optimized for Fluorescence Microplate Reader*
|Shipping||Standard overnight for United States, inquire for international|
|H-phrase||H303, H313, H333|
|Intended use||Research Use Only (RUO)|
|R-phrase||R20, R21, R22|
|Cell Meter™ Cell Viability Assay Kit *Red Fluorescence*|
|Cell Meter™ Cell Viability Assay Kit *Blue Fluorescence with 405 nm Excitation*|
|Cell Meter™ Cell Viability Assay Kit *Blue Fluorescence*|
|Cell Meter™ Cell Viability Assay Kit *Green Fluorescence*|
Fluorescence microplate reader
|Recommended plate||Black wall/clear bottom|
|Instrument specification(s)||Bottom read mode|
AT A GLANCE
- Prepare cells with test compounds
- Remove the medium
- Add CytoCalcein™ NIR working solution (100 µL/well/96-well plate or 25 µL/well/384-well plate)
- Incubate at room temperature or 37°C for 1 hr
- Read fluorescence intensity (bottom read mode) at Ex/Em = 635/670 nm (Cutoff = 665 nm)
Thaw one of each kit component at room temperature before starting the experiment.
PREPARATION OF STOCK SOLUTION
1. CytoCalcein™ NIR stock solution:
Add 20 µL of DMSO (Component B) into the vial of CytoCalcein™ NIR (Component A) and mix them well to make CytoCalcein™ NIR stock solution. Note: 20 µL of CytoCalcein™ NIR stock solution is enough for one plate. Protect from light. For storage, seal tubes tightly.
PREPARATION OF WORKING SOLUTION
Add 20 µL of CytoCalcein™ NIR stock solution into the bottle of Assay Buffer (10 mL, Component C) and mix well to make CytoCalcein™ NIR working solution. Note: This CytoCalcein™ NIR working solution is for one cell plate and stable for at least 2 hours at room temperature.
SAMPLE EXPERIMENTAL PROTOCOL
Plate 100 to 100,000×10 cells per well in a tissue culture microplate with black wall and clear bottom. Add test compounds into the cells and incubate for a desired period of time (such as 24, 48 or 96 hours) in a 37 °C, 5% CO2 incubator.
For blank wells (medium without the cells), add the same amount of compound buffer. The suggested total volume is 100 µL/well/96-well plate, and 25 µL/well/384-well plate. Note: Each cell line should be evaluated on the individual basis to determine the optimal cell density for proliferation or cytotoxicity induction. For proliferation assays, use fewer cells; for cytotoxicity assays, use more cells to start with.
- Treat cells with test compounds as desired. Note: It is not necessary to wash cells before adding compound. However, if tested compounds are serum sensitive, growth medium and serum factors can be aspirated away before adding compounds. Add 100 µL/well (96-well plate) and 25 µL/well (384-well plate) of 1X Hank’s salt solution and 20 mM Hepes buffer (HHBS) or the buffer of your choice after aspiration. Alternatively, cells can be grown in serum-free media.
- Remove the medium from the cells. Note: Medium must be removed before loading CytoCalcein™ NIR working solution.
- Add 100 µL/well (96-well plate) or 25 µL/well (384-well plate) of CytoCalcein™ NIR working solution.
- Incubate plate at room temperature or 37°C for 1 hour, protected from light. Note: The appropriate incubation time depends on the individual cell type and cell concentration used. Optimize the incubation time for each experiment. For non-adherent cells, it is recommended to centrifuge cell plates at 800 rpm for 2 minutes with brake off after incubation.
- Monitor the fluorescence intensity with a fluorescence plate reader (bottom read mode) at Ex/Em = 635/670 nm (Cutoff = 665 nm).
Authors: Li, Jingcheng and Zhang, Jianxiong and Wang, Meng and Pan, Junxia and Chen, Xiaowei and Liao, Xiang
Journal: Biomedical Optics Express (2017): 2599--2610
Authors: Wang, Jingjing and Fa, Jingjing and Wang, Pengyun and Jia, Xinzhen and Peng, Huixin and Chen, Jing and Wang, Yifan and Wang, Chenhui and Chen, Qiuyun and Tu, Xin and others, undefined
Journal: Cellular Signalling (2017)
Authors: Bogert, Nicolai V and Werner, Isabella and Kornberger, Angela and Meybohm, Patrick and Moritz, Anton and Keller, Till and Stock, Ulrich A and Beiras-Fern, undefined and ez, Andres
Journal: Scientific reports (2016)
Authors: Alharbi, Hattan A and Saunders, David MV and Al-Mousa, Ahmed and Alcorn, Jane and Pereira, Alberto S and Martin, Jonathan W and Giesy, John P and Wiseman, Steve B
Journal: Aquatic Toxicology (2016): 81--88
Authors: Brett, Marie-Elena and Crampton, Alex and ra L , undefined and Wood, David K
Journal: Technology (2016): 1--8
Authors: Alharbi, Hattan A and Alcorn, Jane and Al-Mousa, Ahmed and Giesy, John P and Wiseman, Steve B
Journal: Journal of Applied Toxicology (2016)
Authors: Thiebes, Anja Lena and Reddemann, Manuel Armin and Palmer, Johannes and Kneer, Reinhold and Jockenhoevel, Stefan and Cornelissen, Christian Gabriel
Journal: Tissue Engineering Part C: Methods (2016): 322--331
Authors: Liu, Peixi and Zhou, Yingjie and An, Qingzhu and Song, Yaying and Chen, Xi and Yang, Guo-Yuan and Zhu, Wei
Journal: MEDICINE (2016): 1--8
Authors: Thiebes, Anja Lena and Albers, Stefanie and Klopsch, Christian and Jockenhoevel, Stefan and Cornelissen, Christian G
Journal: BioResearch open access (2015): 278--287
Authors: Le Bihan, Olivier and Decossas, Marion and Gontier, Etienne and Gerbod-Giannone, Marie-Christine and Lambert, Olivier
Journal: Journal of structural biology (2015): 470--477
Authors: Li, Deng and Xiang, Xiaocong and Yang, Fei and Xiao, Dongqin and Liu, Kang and Chen, Zhu and Zhang, Ruolan and Feng, Gang
Journal: Biochemical and Biophysical Research Communications (2017)
Authors: Zibek S, Stett A, Koltay P, Hu M, Zengerle R, Nisch W, Stelzle M.
Journal: Biophys J. (2006)
Authors: Klesius PH, Evans JJ, Shoemaker CA, Pasnik DJ.
Journal: Fish Shellfish Immunol (2006): 20
Authors: Bratosin D, Mitrofan L, Palii C, Estaquier J, Montreuil J.
Journal: Cytometry A (2005): 78
Authors: Schoonen WG, Westerink WM, de Roos JA, Debiton E.
Journal: Toxicol In Vitro (2005): 505
Authors: Iwanowicz LR, Densmore CL, Ottinger CA.
Journal: Fish Shellfish Immunol (2004): 127
Authors: Uggeri J, Gatti R, Belletti S, Sc and roglio R, Corradini R, Rotoli BM, Orl and ini G., undefined
Journal: Histochem Cell Biol (2004): 499
Authors: Mueller H, Kassack MU, Wiese M.
Journal: J Biomol Screen (2004): 506
Authors: Goto T, Kajiwara H, Yoshinari M, Fukuhara E, Kobayashi S, Tanaka T.
Journal: Biomaterials (2003): 3885
Authors: Tokudome Y, Sugibayashi K.
Journal: Biol Pharm Bull (2003): 1508