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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. There are a few reagents or assay kits available for quantitating thiol content in biological systems. All the commercial kits either lack sensitivity or have tedious protocols.
ab112158 provides an ultrasensitive fluorometric assay to quantitate thiol content that exists either in a small molecule or on a protein. The proprietary non-fluorescent dye used in the kit becomes strongly fluorescent upon reacting with thiol.
ab112158 can detect as little as 1 picomole of cysteine or GSH in a 100 µL assay volume (10 nM). The assay can be performed in a convenient 96-well or 384-well microtiter-plate format and easily adapted to automation without a separation step. The thiol sensor used in the kit generates a strongly fluorescent adduct upon reacting with a thiol compound. This adduct has spectral properties almost identical to those of fluorescein. In addition, both absorption and emission spectra of the thiol adduct are pH-independent, making ab112158 highly robust. The signal can be easily read by a fluorescence microplate reader at Ex/Em = 490/520 nm.
Please note it is not possible to do a quantification in comparison to the standard.
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ab112158 should be stored desiccated.
|Assay Buffer||1 x 25ml|
|DMSO||1 x 200µl|
|GSH Standard||1 x 62µg|
|Thiol Green Indicator||1 vial|
Our Abpromise guarantee covers the use of ab112158 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Functional Studies||Use at an assay dependent concentration.|
Standard curve: mean of duplicates (+/- SD) with background reads subtracted
GSH measured in tissue lysates showing quantity (nmol) per mg of extracted protein.
Protein concentration for samples varied from 6 mg/mL to 16 mg/mL. Samples were diluted 10-1000 fold.
GSH measured in cell lysates showing quantity (nmol) per 1 mln cells.
Samples with the concentration of 1e8 cells/mL were used. Samples were diluted 100-1000 fold.
GSH measured in biological fluids. Samples were diluted 10-1000 fold.
ab112158 has not yet been referenced specifically in any publications.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"