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 | ![]() ![]() |
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
![The decrease in the fluorescence intensity of ThiolTrace™ Violet 500 with the addition of Campothecin in HL-60 cells. HL-60 cells were treated for 6 hours without (Red line) or with 20 μM Campothecin (Green line) in a 37 <sup>o</sup>C, 5% CO<sub>2</sub> incubator, and then stained with ThiolTrace™ Violet 500 for 20 minutes. The fluorescence intensity was measured using ACEA NovoCyte 3000 flow cytometer with Pacific Orange channel.](/_next/image?url=https%3A%2F%2Fimages.aatbio.com%2Fproducts%2Ffigures-and-data%2Fcell-meter-intracellular-gsh-assay-kit-optimized-for-flow-cytometry-with-405-nm-excitation%2Ffigure-for-cell-meter-intracellular-gsh-assay-kit-optimized-for-flow-cytometry-with-405-nm-excitation_Ly4Rf.jpg&w=3840&q=75)
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: Muri, J., Thut, H., Heer, S., Krueger, C. C., Bornkamm, G. W., Bachmann, M. F., Kopf, M.
Journal: Eur J Immunol (2019): se name="22809.enl" path="C:\Users\aatbi\Drop
Authors: Saisawang, C., Wongsantichon, J., Robinson, R. C., Ketterman, A. J.
Journal: Proteins (2019): se name="22809.enl" path="C:\Users\aatbi\Drop
Authors: Zhang, H., Chen, J., Yang, Y., Wang, L., Li, Z., Qiu, H.
Journal: Anal Chem (2019): 5004-5010
Authors: Vasireddi, M., Crum, A., May, H., Katz, D., Hilliard, J.
Journal: Antiviral Res (2019): 46-52
Authors: Zhang, J., An, H., Ni, K., Chen, B., Li, H., Li, Y., Sheng, G., Zhou, C., Xie, M., Chen, S., Zhou, T., Yang, G., Chen, X., Wu, G., Jin, S., Li, M.
Journal: Cell Death Dis (2019): 321
Authors: Ren, L., Qi, K., Zhang, L., Bai, Z., Ren, C., Xu, X., Zhang, Z., Li, X.
Journal: Biol Trace Elem Res (2019): se name="22809.enl" path="C:\Users\aatbi\Drop
Authors: Chen, H., Wang, X., Gou, S.
Journal: J Inorg Biochem (2019): 133-142
Authors: Khlebnikov, A. I., Schepetkin, I. A., Kishkentaeva, A. S., Shaimerdenova, Z. R., Atazhanova, G. A., Adekenov, S. M., Kirpotina, L. N., Quinn, M. T.
Journal: Molecules (2019): se name="22809.enl" path="C:\Users\aatbi\Drop
Authors: Mlakar, V., Jurkovic Mlakar, S., Lesne, L., Marino, D., Rathi, K. S., Maris, J. M., Ansari, M., Gumy-Pause, F.
Journal: J Exp Clin Cancer Res (2019): 69
Application notes
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Dopamine-Mediated Oxidation of Methionine 127 in α-Synuclein