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Intracellular Glutathione Assay Kit

Pricing & Availability
Regulatory Status
RUO
Other Names
GSH assay kit, glutathione sensor, GSH probe, GSH Live Cell Assay, Glutathione Kit
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Product Citations
publications
a. /Intracellular-Glutathione-Assay-Kit_1_080125
Live cell fluorescence imaging of HeLa cells. HeLa cells were left untreated (left) or treated with 25 µM Menadione (right) for 30 minutes. Menadione induces the formation of reactive oxygen species and depletion of GSH-mediated apoptosis. After treatment, cells were probed using the Intracellular Glutathione Assay Kit. In cells treated with Menadione, green fluorescence is reduced indicating a decrease in intracellular levels of reduced Glutathione. Images were captured using a 40X objective. Scale bar: 20 µm
  • a. /Intracellular-Glutathione-Assay-Kit_1_080125
    Live cell fluorescence imaging of HeLa cells. HeLa cells were left untreated (left) or treated with 25 µM Menadione (right) for 30 minutes. Menadione induces the formation of reactive oxygen species and depletion of GSH-mediated apoptosis. After treatment, cells were probed using the Intracellular Glutathione Assay Kit. In cells treated with Menadione, green fluorescence is reduced indicating a decrease in intracellular levels of reduced Glutathione. Images were captured using a 40X objective. Scale bar: 20 µm
  • b. /Intracellular-Glutathione-Assay-Kit_2_080125
    Jurkat cells were stained using the Intracellular Glutathione Assay Kit (purple histogram) or left unstained (dashed histogram). Dead cells (light gray histogram) have low levels of GSH and low fluorescence intensity.
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421936 1 kit 340€
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Description

The antioxidant reduced glutathione (GSH) plays an essential role in preserving a reduced intracellular environment, and protects cellular macromolecules like DNA, proteins and lipids against oxidizing agents. Reactive oxygen species can oxidize cellular GSH, leading to the loss of intracellular redox homeostasis and activation of the apoptotic signaling cascade.

The Intracellular Glutathione Assay Kit is designed to measure GSH in live cells by flow cytometry and live-cell imaging. This fluorometric assay is based on the detection of GSH in cells using the GSH Probe Green, which is non-fluorescent in its native state but becomes strongly fluorescent upon reacting with glutathione. Fluorescence intensity decreases in cells with low GSH levels, as well as in cells undergoing apoptosis.

This kit provides enough reagents for 100 tests (100 mL total working solution for up to 1x108 cells). A single test is defined as 1 mL of working solution for up to 1x106 cells.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human
Formulation
• 1 vial (50 μL) of Glutathione Probe Green
• 1 bottle (100 mL) of Glutathione Assay Buffer
• 1 vial (500 μL) of DMSO
Storage & Handling
The kit should be stored at < -15°C protected from light.
Application

Live cell imaging - Quality tested
FC - Verified
Application Notes
Detection/Imaging Guidelines:
- Fluorescence microscope filter set: GFP/FITC filter set
- Flow cytometry channels:
  • Ex/Em = 590/620 nm
  • Excitation: 488 nm laser
  • Emission: 530/30 nm filter
  • Instrument specification(s): FITC channel

Components and storage
This kit is stored at -20°C. For convenience, the buffer and DMSO can be stored at 4°C.
- 1 vial of Glutathione Probe Green (50 μL) (store at -20°C and minimize light exposure)
- 1 bottle of Glutathione Assay Buffer (100 mL)
- 1 vial of DMSO (500 μL)

Required Materials Not Included:
- For live cell imaging: tissue culture vessel with coverslip bottom

Assay Protocol:
This kit provides enough reagents for 100 tests (for up to 1x108 cells). A single test is defined as 1 mL of Glutathione Probe Green working solution for up to 1x106 cells.

Preparation of 200X Glutathione Probe Green: 
Bring DMSO and Glutathione Probe Green provided in the kit to room temperature. Add 500 µL of DMSO into the vial containing 50 µL of Glutathione Probe Green Stock Solution and mix well to make 200X Glutathione Probe Green. If not using immediately, aliquot the 200X Glutathione Probe Green and store at -20°C protected from light. Avoid freeze thaw cycles.
 

Experimental Protocol:
1. Bring all components of the kit to room temperature before starting. Treat your cell samples under desired experimental conditions in complete media. 
2. Prepare the Glutathione Probe Green Working Solution by adding 5 µL of the 200X Glutathione Probe Green to 1 mL media for up to 1x106 cells. For larger cell amounts, scale up accordingly. 
3. Gently remove media from your cells and add the Glutathione Probe Green Working Solution and incubate the samples at 37°C for 15-30 minutes. 
4. For live cell imaging: Gently wash the cells 3 times with Glutathione Assay Buffer or buffer of choice  and add the desired volume of Glutathione Assay Buffer or your buffer of choice for imaging. Observe cells with an inverted fluorescence microscope using the GFP/FITC filter set.

For flow cytometry: Spin down cells and resuspend in the desired volume of Glutathione Assay Buffer or your buffer of choice. Monitor the fluorescence intensity on a flow cytometer using the FITC (530/30 nm) channel.

Antigen Details

Distribution

Cytoplasm

Function
Reduced glutathione sensor
Biology Area
Cell Proliferation and Viability, Mitochondrial Function
Antigen References
  1. Maulucci G, et al. 2009. Biosens Bioelectron. 25:682-7.
  2. Bellomo G, et al. 1997. Microsc Res Tech. 36:243-52.
  3. Baylor KJ, et al. 1997. Biochem Soc Trans. 25:47S.
  4. Jin T, et al. 2008. Int J Mol Sci: 9:2044-2061.
Gene ID
NA

Related FAQs

There are no FAQs for this product.
Go To Top Version: 1    Revision Date: 08/01/2025

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

BioLegend, the BioLegend logo, and all other trademarks are property of BioLegend, Inc. or their respective owners, and all rights are reserved.

 

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Toll-Free Phone: 1-877-Bio-Legend (246-5343) Phone: (858) 768-5800 Fax: (877) 455-9587

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