Free Thiol Assay Kit (Fluorometric)
4
(1 Review)
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(6 Publications)
- FuncS
Lab
Functional Studies - Free Thiol Assay Kit (Fluorometric) (AB112158)
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.
- FuncS
Lab
Functional Studies - Free Thiol Assay Kit (Fluorometric) (AB112158)
GSH measured in biological fluids. Samples were diluted 10-1000 fold.
- FuncS
Supplier Data
Functional Studies - Free Thiol Assay Kit (Fluorometric) (AB112158)
GSH and Cysteine dose responses were measured in a 96-well black plate with ab112158 using a microplate reader. As low as 10 nM (1 pmol/well) of GSH or Cysteine can be detected with 10 minutes incubation time (n=3). The insert shows the low levels of thiol detection.
- FuncS
Lab
Functional Studies - Free Thiol Assay Kit (Fluorometric) (AB112158)
Standard curve : mean of duplicates (+/- SD) with background reads subtracted
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Product details
Free Thiol Assay Kit (Fluorometric) (ab112158) provides an ultrasensitive fluorometric assay to quantitate thiol content that exists in small molecules such as free cysteine, glutathione, and cysteine residues.
This product is not suitable for assaying thiols incorporated within proteins. It is designed for thiols in free small molecules like amino acids.
The assay is based on a proprietary non-fluorescent dye tha generates a strongly fluorescent adduct upon reacting with a thiol compound. The signal produced by the adduct can be easily read by a fluorescence microplate reader at Ex/Em = 490/520 nm. In addition, both absorption and emission spectra of the thiol adduct are pH-independent, making the assay highly robust. There are few reagents or assay kits available for quantitating thiol content in biological systems, and the majority of commercial kits either lack sensitivity or have tedious protocols. The assay can detect as little as 1 picomole of cysteine or GSH in a 100 uL assay volume (10 nM).
Please note it is not possible to do a quantification in comparison to the standard.
Free Thiol assay protocol summary:
- add samples and standards to wells
- add reaction mix
- analyze with microplate reader for 10 min to 1 hr
Other Notes
This product was previously called Thiol Quantification Assay Kit (Fluorometric).
This product detects thiol groups on small molecules. If you want to detect thiol groups in proteins, we recommend Total Protein Thiol Quantification Assay Kit (Colorimetric) (ab219272).
What's included?
Properties and storage information
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Storage information
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
Thiol groups in cysteine residues participate in forming disulfide bridges which are critical for maintaining protein structure and stability. These links can affect the protein's shape and function and they change based on the cellular environment. Some proteins possess dithiol groups due to the presence of multiple thiol pairs making them active in catalytic functions. These proteins may be part of larger complexes like enzymes that catalyze redox reactions influencing processes such as cellular respiration and detoxification pathways.
Pathways
Thiol-containing proteins play significant roles in redox balance and signal transduction. These proteins participate in pathways such as the oxidative stress response and cellular signaling cascades. For example glutathione a tripeptide with a thiol group is critical in the antioxidant pathway protecting cells from oxidative damage with enzymes like glutathione peroxidase. Moreover thiol groups can modulate the function of transcription factors impacting gene expression pathways by altering DNA and protein interactions.
Product protocols
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Publications (6)
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Frontiers in ophthalmology 3:1274825 PubMed38983035
2024
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iScience 26:108566 PubMed38144458
2023
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Aging cell 22:e13797 PubMed36823285
2023
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Cell chemical biology 28:463-474.e7 PubMed33357462
2020
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Scientific reports 10:2140 PubMed32034177
2020
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Scientific reports 9:9186 PubMed31235767
2019
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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