Name: Amplite® Rapid Fluorimetric Total Thiol Quantitation Assay Kit *Green Fluorescence*
Brand: AAT Bioquest
SKU: 5528
Price: 237 USD
Availability: InStock

Amplite® Rapid Fluorimetric Total Thiol Quantitation Assay Kit

Green Fluorescence

Amplite® Rapid Fluorimetric Total Thiol Quantitation Assay Kit is a fluorimetric assay kit for quantification of free thiols in small molecules and macromolecules such as proteins.

Detection Principle: Uses a nonfluorescent thiol-reactive dye that becomes strongly fluorescent after reaction with free thiols.
Sensitivity: Detects as little as 1 pmol of cysteine or GSH in a 100 µL assay volume (10 nM).
Workflow: Provides a rapid, one-step assay that can be readily adapted to automation.
Instrument Compatibility: Supports 96- or 384-well formats with fluorescence detection at Ex/Em 490/525 nm.

GSH and cysteine dose responses were measured on a solid black 96-well plate with Amplite® Fluorimetric Thiol Quantitation Assay Kit using a NOVOstar microplate reader (BMG Labtech).

Protocol

SDS

COA

Catalog	Size	Price	Quantity
5528	200 Tests	Price

Overview

The detection and measurement of free thiol (such as free cysteine, glutathione and cysteine residues in proteins) is one of the essential tasks for investigating biological processes and events in many biological systems. The monitoring of reduced (GSH) and oxidized glutathione in biological samples is essential for evaluating the redox and detoxification status of cells and tissues in relation to the protective role of glutathione against oxidative and free-radical-mediated cell injury. Disorders of cysteine metabolism include cystinosis, an autosomal recessive disease produced by a defect in lysosomal transport, and cystinuria, a common heritable disorder of amino acid transport. There are few reagents or assay kits available for quantitating thiols in biological systems. However, all the commercial kits either lack sensitivity or have tedious protocols. Our Amplite® Fluorimetric Thiol Quantitation Kit provides an ultrasensitive fluorimetric assay for quantitating thiols that exist either in a small molecule or a macromolecule such as protein. The kit uses a proprietary water-soluble non-fluorescent dye that becomes strongly fluorescent upon reacting with thiol. The kit provides a sensitive, one-step fluorimetric method to detect as little as 1 picomole of cysteine or GSH in a 100 µL assay volume (10 nM in concentration). The assay is rapid and robust. It can be performed in a convenient 96-well or 384-well microtiter-plate format and easily adapted to automation. For rapid quantifying thiol groups in a protein, we recommend you use our kit #5529 that is optimized for quantifying protein thiols.

References

View all 26 references: Citation Explorer

Covalent surface chemistry gradients for presenting bioactive peptides

Authors:

Kipper MJ, Kleinman HK, Wang FW.

Journal:

Anal Biochem (2007): 175

Dansyl glutathione as a trapping agent for the quantitative estimation and identification of reactive metabolites

Authors:

Gan J, Harper TW, Hsueh MM, Qu Q, Humphreys WG.

Journal:

Chem Res Toxicol (2005): 896

Gold nanoparticle-based detection of genomic DNA targets on microarrays using a novel optical detection system

Authors:

Storhoff JJ, Marla SS, Bao P, Hagenow S, Mehta H, Lucas A, Garimella V, Patno T, Buckingham W, Cork W, Muller UR.

Journal:

Biosens Bioelectron (2004): 875

Fluorometric polyethyleneglycol-peptide hybrid substrates for quantitative assay of protein disulfide isomerase

Authors:

Christiansen C, St Hilaire PM, Winther JR.

Journal:

Anal Biochem (2004): 148

Assay of total homocysteine and other thiols by capillary electrophoresis and laser-induced fluorescence detection. II. Pre-analytical and analytical conditions

Authors:

Bayle C, Issac C, Salvayre R, Couderc F, Causse E.

Journal:

J Chromatogr A (2002): 255

Usage and storage

Intended use

Research Use Only (RUO)

Instrument settings

Fluorescence microplate reader
Excitation	490 nm
Emission	525 nm
Cutoff	515 nm
Recommended plate	Solid black

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Page updated on October 28, 2026

Component A: Thiolite™ Green 520WS	1 vial
Component B: Assay Buffer	1 bottle (25 mL)
Component C: GSH Standard	1 vial (62 µg)