Glutathione Colorimetric Assay kit

Detects either total glutathione or exclusively GSH

Glutathione (GSH) is the major intracellular low-molecular-weight thiol that plays a critical role in the cellular

defense against oxidative stress in mammalian cells. HansaBioMed’s ApoGSH™ Glutathione Colorimetric

Assay Kit provides a convenient, colorimetric method for analyzing either total glutathione or the reduced form

glutathione alone using a microtiter plate reader. The assay is based on the glutathione recycling system by

DTNB and glutathione reductase (Fig. 1). DTNB and glutathione (GSH) react to generate 2-nitro-5-thiobenzoic

acid which has yellow color. Therefore, GSH concentration can be determined by measuring absorbance at

412 nm. The generated GSSG can be reduced back to GSH by glutathione reductase, and GSH reacts with

DTNB again to produce more 2-nitro-5-thiobenzoic acid. Therefore, the recycling system dramatically

improves the sensitivity of total glutathione detection. The kit includes the 5-Sulfosalicylic acid (SSA) for the

removal of proteins from samples and for the protection of GSH oxidation and γ-glutamyl transpeptidase

reaction. The kit can quantify glutathione from 1-100 ng/well in a 200 l reaction. For detecting lower

glutathione concentrations, such as in blood samples, increasing reaction time will generate stronger signal.

The kit can also specifically detect the reduced form of glutathione (GSH) by omitting the glutathione

reductase from the reaction mixture. The sensitivity for detecting the reduced form of glutathione (without

recycling system) is 100 times lower than detecting the total glutahione.

Features

• Simple procedure; takes only ~2-3 hours

• Fast and convenient

Application & Specifications

• The Colorimetric Glutathione Detection Kit provides a simple in vitro assay for detecting the GSH changes in

  apoptosis and other pathological processes

• Detection method- Absorbance (412 nm)

• Sample type- Cell and Tissue lysates, culture media, urine, plasma and serum, as well as many other

  biological fluids

• Species reactivity- Mammalian

Kit Components

• Glutathione Reaction Buffer

• Glutathione Substrate (DTNB)

• NADPH Generating Mix (lyophilized)

• Glutathione Reductase (lyophilized)

• Sulfosalicylic Acid (SSA, 1 gram)

• GSH Standard (lyophilized, MW 307)

- Storage Conditions: -20°C

- Shipping Conditions: gel pack

- Usage: For research use only.  Not for use in humans

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