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GSH/GSSG-Glo™ Assay

A Luminescent-Based Assay to Detect and Quantify Total Glutathione Ratios

  • Measures physiologically relevant GSH/GSSG ratios
  • Simple protocol minimizes sample handling, reducing variability
  • Stable assay signal amenable to high throughput analysis

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Catalog number selected: V6611

$ 518.00
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GSH/GSSG-Glo™ Assay
10ml
$ 518.00
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Valuable Measurement of Glutathione (GSH-to-GSSG) Ratios

The GSH/GSSG-Glo™ Assay is a luminescence-based system that detects and quantifies total glutathione (GSH + GSSG), GSSG and GSH-to-GSSG ratios in cultured cells. Reduced glutathione (GSH) and oxidized glutathione (GSSG) both exist in healthy cells, the majority of which is in reduced (GSH) form. A change in GSH and GSSG levels are important when assessing toxicological responses and an indicator of oxidative stress, potentially leading to apoptosis or cell death. The GSH and GSSG detection assay provides a simple, rapid multiwell-plate format where stable luminescent signals are correlated with either the total GSH or the GSSG concentration of a sample directly in culture wells.

Both total glutathione and GSSG determinations are based on the reaction where GSH-dependent conversion of a GSH probe, Luciferin-NT, to luciferin by a glutathione-S-transferase enzyme is coupled to a firefly luciferase reaction. Light from luciferase is dependent on the amount of luciferin formed, which is in turn dependent on the amount of GSH present. This makes the luminescent signal proportional to the amount of GSH. Determining the total glutathione and amount of GSSG are performed in parallel reactions. In one configuration, the assay reagents measure total glutathione using a reducing agent that converts all the glutathione, GSH and GSSG in a cell lysate to the reduced form, GSH. In a second configuration, the assay reagents are used to measure only the oxidized form, GSSG. In this case, a reagent is added that blocks all the GSH while leaving the GSSG intact. This blocking step is followed by a reducing step that converts the GSSG to GSH for quantification in the luminescent reaction. Because the assays are performed directly on cells in culture wells, loss of GSH or GSSG is minimized, reducing variability.

GSH/GSSG Ratio Determination in Different Cell Lines

Cell Line Cells/Well Ratio GSH/GSSG
Vehicle Menadione
HeLa 5,000 63.8 2.2
HepG2 10,000 92.6 2.2
HepaRG® 50,000 35.3 2.5
A549 5,000 87.9 10.9
Rat hepatocytes 50,000 19.8 1.1

The GSH/GSSG-Glo™ Assay. GSH-dependent conversion of a GSH probe, Luciferin-NT, to luciferin by a glutathione S-transferase enzyme is coupled to a firefly luciferase reaction.

Limit of Detection Using Reduced (GSH) and Oxidized (GSSG) Glutathione

GSH (Panel A) and GSSG (Panel B) standards were assayed in a 96-well plate using the GSH/GSSG-Glo™ Assay and a GloMax® Luminometer. The results were averaged and plotted on the graphs above. A linear trend line was generated and correlation coefficients calculated for the fit of the data to the trend line. Limits of detection were 2.5nM GSH and 0.5nM GSSG.

Specifications

You are viewing: V6611 Change Configuration

What's in the box?

Item Part # SizeConcentrationAvailable Separately

Passive Lysis Buffer, 5X

E194B 1 × 1ml View Product

DTT

P117B 1 × 250μl100mM View Product

Reconstitution Buffer with Esterase

V144A 1 × 10ml

Glutathione Reaction Buffer

V163A 1 × 10ml

NEM, 25mM

V164A 1 × 50μl

Luciferin-NT

V687A 1 × 100μl

Glutathione S-Transferase

V689B 1 × 500μl

Glutathione, 5mM

V690A 1 × 100μl

Luciferin Detection Reagent

V859A 1 × 1 each

SDS

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Download SDSPDF (323 KB) – English (United States)

Certificate of Analysis

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Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

BB

Patents and Disclaimers

European Pat. No. 1131441 and Japanese Pat. No. 4520084.

Certain applications of this product may require licenses from others.

Specifications

You are viewing: V6612 Change Configuration

What's in the box?

Item Part # SizeConcentrationAvailable Separately

Passive Lysis Buffer, 5X

E194C 1 × 5ml View Product

DTT

P117C 1 × 1.25ml100mM View Product

Reconstitution Buffer with Esterase

V144B 1 × 50ml

Glutathione Reaction Buffer

V163B 1 × 50ml

NEM, 25mM

V164B 1 × 250μl

Luciferin-NT

V687B 1 × 500μl

Glutathione S-Transferase

V689B 3 × 500μl

Glutathione, 5mM

V690A 2 × 100μl

Luciferin Detection Reagent

V859B 1 × 1 each

SDS

Choose language:

Download SDSPDF (323 KB) – English (United States)

Certificate of Analysis

See all certificates for V6612

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

BB

Patents and Disclaimers

European Pat. No. 1131441 and Japanese Pat. No. 4520084.

Certain applications of this product may require licenses from others.

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