Cell Meter™ Intracellular GSH Assay Kit *Optimized for Flow Cytometry with 405 nm excitation*
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Telephone | 1-800-990-8053 |
Fax | 1-800-609-2943 |
sales@aatbio.com | |
International | See distributors |
Bulk request | Inquire |
Custom size | Inquire |
Shipping | Standard overnight for United States, inquire for international |
Excitation (nm) | 415 |
Emission (nm) | 499 |
H-phrase | H303, H313, H333 |
Hazard symbol | XN |
Intended use | Research Use Only (RUO) |
R-phrase | R20, R21, R22 |
UNSPSC | 12352200 |
Cell Meter™ Intracellular GSH Assay Kit *Optimized for Flow Cytometry* |
Overview | SDSProtocol |
Excitation (nm) 415 | Emission (nm) 499 |
Platform
Flow cytometer
Excitation | 405 nm laser |
Emission | 525/50 nm filter |
Instrument specification(s) | Pacific Orange channel |
Components
Example protocol
AT A GLANCE
Protocol summary
- Prepare cells with test compounds at a density of 5 × 105 to 1 × 106 cells/mL
- Prepare and add ThiolTraceTM Violet 500 working solution to cells
- Incubate at 37oC for 20 to 30 minutes
- Read fluorescence intensity at Ex/Em = 405/525 nm-Pacific Orange filter set
Important notes
Thaw at room temperature before starting the experiment.
PREPARATION OF STOCK SOLUTION
ThiolTraceTM Violet 500 stock solution (500X):
Add 200 µL of DMSO (Component C) into the vial of ThiolTraceTM Violet 500, and mix well. Note: Aliquot and stored the unused ThiolTraceTM Violet 500 stock solution at -20 oC. Avoid repeated freeze/thaw cycles.
PREPARATION OF WORKING SOLUTION
ThiolTraceTM Violet 500 working solution (1X):
Add 2 µL of ThiolTraceTM Violet 500 stock solution into 1 mL of Assay Buffer (Component B) or buffer of your choice, and mix well. Note: ThiolTraceTM Violet 500 working solution can be prepared in the cell culture medium containing serum.
For guidelines on cell sample preparation, please visit
https://www.aatbio.com/resources/guides/cell-sample-preparation.html
SAMPLE EXPERIMENTAL PROTOCOL
- Treat cells with test compounds for a desired period of time. Note: For adherent cells, gently lift the cells with 0.5 mM EDTA to keep the cells intact, and wash the cells once with serum-containing media prior to the incubation with ThiolTrace™ Violet working solution. Note: The appropriate incubation time depends on the individual cell type and cell concentration used. Optimize the incubation time for each experiment.
- Centrifuge the cells at 1000 rpm for 4 minutes, and wash cells in 1 mL of buffer of your choice (Optional).
- Resuspend cells in 1 mL ThiolTraceTM Violet 500 working solution and incubate them at 37oC incubator for 20 to 30 minutes.
- Monitor the fluorescence intensity with a flow cytometer using Pacific Orange filter set (Ex/Em = 405/525 nm). Gate on the cells of interest, excluding debris.
Images
Citations
Authors: Ogino, S., Konishi, H., Ichikawa, D., Matsubara, D., Shoda, K., Arita, T., Kosuga, T., Komatsu, S., Shiozaki, A., Okamoto, K., Kishimoto, M., Otsuji, E.
Journal: Cancer Sci (2019): 795-804
Authors: Husarcikova, J., Schallmey, A.
Journal: J Biotechnol (2019): 1-7
Authors: Dong, X., Yang, Y., Zhou, Y., Bi, X., Zhao, N., Zhang, Z., Li, L., Hang, Q., Zhang, R., Chen, D., Cao, P., Yin, Z., Luo, L.
Journal: Cell Death Differ (2019): se name="22809.enl" path="C:\Users\aatbi\Drop
Authors: Wang, F. X., Liang, J. H., Zhang, H., Wang, Z. H., Wan, Q., Tan, C. P., Ji, L. N., Mao, Z. W.
Journal: ACS Appl Mater Interfaces (2019): 13123-13133
Authors: He, N., Bai, S., Huang, Y., Xing, Y., Chen, L., Yu, F., Lv, C.
Journal: Anal Chem (2019): 5424-5432
Authors: Wang, Q., Zhang, Y., Wang, X., Wu, Y., Dong, C., Shuang, S.
Journal: Analyst (2019): 1988-1994
Authors: Sahebekhtiari, N., Fern and ez-Guerra, P., Nochi, Z., Carlsen, J., Bross, P., Palmfeldt, J.
Journal: Biochim Biophys Acta Mol Basis Dis (2019): 126-135
Authors: Wang, S., Zhang, J., Jun, F., Bai, Z.
Journal: BMC Med Genet (2019): 22
Authors: Kawahara, B., Ramadoss, S., Chaudhuri, G., Janzen, C., Sen, S., Mascharak, P. K.
Journal: J Inorg Biochem (2019): 29-39
Authors: Luethcke, K. R., Ekena, J., Chun, R., Trepanier, L. A.
Journal: J Vet Intern Med (2019): ersion="1.0" encoding="UTF-8" ?>22809.enlEndN
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