Live or Dead™ Cell Viability Assay Kit *Red/Blue Dual Fluorescence*
|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|
|Live or Dead™ Cell Viability Assay Kit *Green/Red Dual Fluorescence*|
|Excitation||Cy5 filter (alive), DAPI filter (dead)|
|Emission||Cy5 filter (alive), DAPI filter (dead)|
|Recommended plate||Black wall/clear bottom|
Fluorescence microplate reader
|Excitation||610, 360 nm|
|Emission||650, 450 nm|
|Cutoff||630, 420 nm|
|Recommended plate||Black wall/clear bottom|
|Instrument specification(s)||Bottom read mode|
AT A GLANCE
- Prepare cells with test compounds
- Add dye-working solution
- Incubate at room temperature or 37°C for 30 minutes to 1 hour
- Monitor fluorescence intensity (bottom read mode) at Ex/Em = 610/650 nm (Cutoff = 630 nm, Red) and Ex/Em = 360/450 nm (Cutoff =420 nm, Blue) or fluorescence microscope with Cy5 channel (live) and DAPI channel (dead)
Thaw all the kit components at room temperature before starting the experiment.
For guidelines on cell sample preparation, please visit:
PREPARATION OF WORKING SOLUTION
Add 5 µL of 200X Cellbrite™ Red (Component A) and 5 µL of 200X Nuclear Blue™ DCS1 (Component C) into 1 mL of Assay Buffer (Component B) and mix well to make dye-working solution. This dye-working solution is stable for at least 1 hour at room temperature.
Note: As the optimal staining conditions may vary depending on different cell types, it’s recommended to determine the appropriate concentration of Component A and C individually.
SAMPLE EXPERIMENTAL PROTOCOL
- Prepare cells according to the standard protocol. Note: We treated HeLa cells with staurosporine (SS) for 4 hours at 37ºC to induce cell apoptosis. See Figure 1 for details.
- Replace growth medium with 100 µL/well (96-well plate) or 25 µL/well (384-well plate) of dye-working solution.
- Incubate the dye-working solution plate at room temperature or 37°C for 30 minutes to 1 hour, protected from light.
- Wash cells with HHBS, PBS or buffer of your choice twice.
- Add 100 µL/well (96-well plate) or 25 µL/well (384-well plate) of Assay Buffer (Component B) into the cells.
- Monitor the fluorescence signal under a fluorescence microscope with Texas Red or Cy5 filter for live cells, and DAPI filter for dead cells. The fluorescence intensity can also be analyzed with a fluorescence microplate reader (bottom read mode) at Ex/Em = 610/650 nm (Cutoff = 630 nm, Red) and Ex/Em = 360/450 nm (Cutoff =420 nm, Blue).
Authors: Soykan, Merve Nur and Altug, Burcugul and Bas, Harun and Ghorbanpoor, Hamed and Avci, Huseyin and Eroglu, Sertac and Sengel, Sultan Butun and Sariboyaci, Ayla Eker and Bagis, Sibel Gunes and Uysal, Onur and others,
Journal: Macromolecular Bioscience (2023): 2300204
Authors: Ozel, Ceren and Apaydin, Elif and Sariboyaci, Ayla Eker and Tamayol, Ali and Avci, Huseyin
Journal: Colloids and Surfaces B: Biointerfaces (2023): 113197
Authors: Misra, Rahul and Rajic, Mathew and Sathiyamoorthy, Krishnan and Karshafian, Raffi
Journal: Journal of Drug Delivery Science and Technology (2020): 102261
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: 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