Intracellular gsh assay kit

Manufactured by Abcam

The Intracellular GSH Assay Kit is a fluorometric assay that allows for the quantification of glutathione (GSH) levels within cells. The kit provides a simple, direct, and sensitive method for measuring intracellular GSH concentrations.

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Lab products found in correlation

H2dcfda, by Thermo Fisher Scientific (2 mentions)

Close spelling variants of this product

GSH Assay Kit (28 citations) GSH kit (4 citations) Intracellular GSH detection assay kit (2 citations) Intracellular glutathione (GSH) Detection Assay Kit (2 citations)

2 protocols using intracellular gsh assay kit

Quantifying Cellular Redox Status

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TSCs under the indicated treatment were transfected with ratiometric redox plasmids pLPCX-Grx1-roGFP2 (used for cytoplasmatic GSH), pLPCX-mito-Grx1-roGFP2 (used for mitochondrial GSH), pLPCX-roGFP2-Orp1 (used for cytoplasmatic H₂O₂) and pLPCX-mito-roGFP2-Orp1(used for mitochondrial H₂O₂) (ref. 48 (link)) and analyzed by FACS. The 405/488 nm ratio was calculated for gated GFP-positive cells.
Cellular ROS content was measured by incubation with 5 μM H2DCF-DA (Thermo Fisher Scientific) for 20 min in phenol red-free medium. The cells were washed in PBS buffer, trypsinized and resuspended in 500 μl PBS.
Total glutathione (GSH) levels were tested using intracellular GSH assay kit (Abcam) following the manufacturer's instructions. Fluorescence intensity was visualized by flow cytometry using FL1 channel.

Castex J., Willmann D., Kanouni T., Arrigoni L., Li Y., Friedrich M., Schleicher M., Wöhrle S., Pearson M., Kraut N., Méret M., Manke T., Metzger E., Schüle R, & Günther T. (2017). Inactivation of Lsd1 triggers senescence in trophoblast stem cells by induction of Sirt4. Cell Death & Disease, 8(2), e2631-.

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Cytokine Production and Tumor Cell Viability Analysis

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For analyzing cytokine production by CD4⁺ T cells, tumor samples were digested and gently dissociated into single-cell suspension. Red blood cells were removed by ACK lysing buffer. After washing with PBS, cells were seeded in 12 well plate and stimulated with PMA and ionomycin for 4 hours. Following stimulation, cells were harvested and stained for surface markers followed by intracellular cytokine staining (ICS). To measure tumor cell viability after culture, cells at ~70% confluence were treated overnight under the specified conditions. Cells were harvested for Annexin V staining following manufacturer’s instruction. Following washing with PBS, cells were resuspended in 400ul 1x binding buffer and labeled with DAPI (0.5ug/ml). DAPI⁺ cells were counted as dead cells, Annexin V⁺DAPI⁻ cells were considered apoptotic cells. The levels of reduced GSH in tumor cells were performed using the Intracellular GSH Assay Kit (Abcam) following the manufacturer’s instruction. DAPI was added to each sample before FACS analysis. Data were acquired using a LSRII and analyzed with FlowJo software (Treestar).

Habtetsion T., Ding Z.C., Pi W., Li T., Lu C., Chen T., Xi C., Spartz H., Liu K., Hao Z., Mivechi N., Huo Y., Blazar B.R., Munn D.H, & Zhou G. (2018). Alteration of Tumor Metabolism by CD4+ T Cells Leads to TNF-α-Dependent Intensification of Oxidative Stress and Tumor Cell Death. Cell metabolism, 28(2), 228-242.e6.

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