Amplite® Rapid Colorimetric Total Protein Thiol Quantitation Assay Kit

Amplite® Rapid Colorimetric Total Protein Thiol Quantitation Assay Kit is a colorimetric assay kit for quantification of free thiol groups in proteins.

Detection Principle: Quantifies protein thiol content from the absorbance of the purified Thiol Blue™ adduct.
Quantitation Method: Determines the thiol-to-protein ratio from absorbance at 680 nm and 280 nm.
Workflow: Uses a single spin-column purification step to remove excess Thiol Blue™ before measurement.
Instrument Compatibility: Compatible with traditional cuvettes, NanoDrop™ spectrophotometers, and 96-well absorbance plate readers using UV-transparent plates.

<p>The Amplite® Rapid Colorimetric Thiol Quantitation Assay Principle</p>

Protocol

SDS

COA

Catalog	Size	Price	Quantity
5529	2 Tests	Price

Overview

Protein thiols are very important to protein structure, protein function and biological system redox environment. For example, albumin is the most abundant protein in plasma and the free thiol present in the albumin protein are considered as major plasma antioxidants in the body. The change of thiol status in albumin is related to a lot of diseases and disorders, such as kidney disease and Parkinson's disease. Although there are a few reagents or assay kits available for quantitating the total thiol content in biological systems, a key challenge is to have a rapid and accurate method to quantify the amount of free thiol group in a specific protein. Amplite® Rapid Colorimetric Thiol Quantitation Kit provides an accurate method to quantify free thiol group using our proprietary thiol sensor, Thiol Blue™, which has the maximum absorbance at ~680 nm. Thiol Blue™ reacts with the protein samples that contain free thiol groups. The resulted thiol adduct is run through a single spin column to remove the excess Thiol Blue™ sensor, and the absorption spectrum of the purified product is measured. The amount of thiol to protein ratio is calculated from the absorbance ratio of 680 nm and 280 nm. This Amplite® Rapid Colorimetric Thiol Quantitation Kit can be performed in a traditional cuvette, NanoDrop™ Spectrophotometer or a convenient 96-well absorbance plate reader with a UV-transparent plate.

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

Spectrophotometer
Absorbance	250 nm~750 nm
Recommended plate	Cuvette

Absorbance microplate reader
Absorbance	280 and 680 nm
Recommended plate	Clear bottom

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Page updated on October 28, 2026

Component A: Thiol Blue™ Sensor	2 vials (One vial is for 50 ~ 100 μg protein)
Component B: Assay Buffer	1 bottle (15 mL)
Component C: Spin Column	2 columns
Component D: Washing Tube (2 mL)	2 tubes
Component E: Collecting Tube (1.5 mL)	2 tubes