- Home
- IVD
- By Technology Types
- By Diseases Types
- By Product Types
- Research
- Resource
- Distributors
- Company
| Product Name | GSH/GSSG Ratio Fluorometric Assay Kit, Thiol-Selective Probe |
| Catalog No. | CMTR-HMM-0076 |
| Description | Fluorometric assay kit for separate quantification of reduced glutathione (GSH) and oxidized glutathione (GSSG) in cell and tissue lysates, enabling calculation of the GSH/GSSG ratio — a key indicator of cellular oxidative stress and redox status. The assay uses a thiol-selective fluorogenic probe that reacts specifically with free thiol groups of GSH to generate a fluorescent product (Ex/Em 490/520 nm). For GSSG measurement, a separate sample aliquot is pretreated with 2-vinylpyridine to derivatize GSH (preventing its detection), then the remaining GSSG is enzymatically reduced to GSH by glutathione reductase with NADPH, enabling total oxidized glutathione quantification. The GSH/GSSG ratio typically ranges from >100:1 in healthy cells to <10:1 in oxidative stress conditions. |
| Intended Use | Quantitative determination of intracellular GSH and GSSG levels and GSH/GSSG ratio as a cellular redox status indicator in oxidative stress, drug toxicity, cancer metabolism, neurodegenerative disease, and aging research. |
| Principle / Technology | Thiol-selective fluorogenic probe reacts with GSH's free -SH group generating fluorescent adduct; GSSG measured after GSH derivatization with 2-vinylpyridine and enzymatic reduction to detectable GSH; ratiometric calculation provides redox balance assessment. |
| Detection Method | Fluorescence microplate reader (Ex/Em 485/530 nm); endpoint measurement after 30-minute incubation at room temperature. |
| Sample Type | Cell lysates (5×10^4-2×10^6 cells); fresh and snap-frozen tissue homogenates; whole blood and plasma (with acid deproteinization). |
| Performance Range / Specifications | GSH linear range: 0.01-1 µM; GSSG linear range: 0.005-0.5 µM; GSH/GSSG ratio measurable from 1:1 to >500:1 with <15% CV. |
| Sensitivity / LOD | GSH detection limit: 2 pmol/well; GSSG detection limit: 1 pmol/well; signal-to-background ratio >20:1 at 50 pmol GSH/well. |
| Specificity | Thiol-reactive probe is specific for free -SH groups; 2-vinylpyridine derivatization specifically blocks GSH; glutathione reductase specifically reduces GSSG; minimal interference from cysteine, homocysteine, or protein thiols in deproteinized samples. |
| Reaction Conditions / Protocol | Deproteinize lysates with 5% metaphosphoric acid (included); neutralize; split into GSH and GSSG aliquots; derivatize GSSG aliquot with 2-vinylpyridine; add probe + enzyme mix; incubate 30 min; measure fluorescence. |
| Components / Formulation | Fluorogenic thiol probe (lyophilized), GSH standard (10 µmol), GSSG standard (5 µmol), glutathione reductase, NADPH, 2-vinylpyridine, metaphosphoric acid (MPA) deproteinizing reagent, assay buffer, GSH quencher, 96-well black plate. |
| Storage Conditions | Store probes, enzymes, and standards at -20 °C; buffer and MPA reagent at 2-8 °C; 2-vinylpyridine at room temperature. |
| Shelf Life | 6 months from date of manufacture. |
| Package Specifications | 100 tests (GSH + GSSG in duplicate), 400 tests. |
| Product Form | Lyophilized probe and enzymes; liquid standards and reagents. |
| Quality Control | Each lot tested for GSH and GSSG standard curve linearity (R² >0.995); probe reactivity verified; GSH recovery 90-110% in spiked cell lysates. |
| Key Features | Separate GSH and GSSG quantification; thiol-specific fluorogenic probe; GSH/GSSG ratio calculation; includes deproteinization reagents; high sensitivity (low pmol range); compatible with tissue and cell samples. |
| Purity | Fluorogenic probe ≥98%; GSH and GSSG standards ≥99% by HPLC; glutathione reductase specific activity ≥200 U/mg. |
| Concentration | GSH standard: 1 mM stock; GSSG standard: 0.5 mM stock. |
| Activity / Unit Definition | Glutathione reductase: 1 unit reduces 1 µmol GSSG per minute at pH 7.5, 25 °C. |
| Molecular Weight | GSH: 307.32 g/mol; GSSG: 612.63 g/mol. |
| Source / Origin | Recombinant yeast glutathione reductase; synthetic fluorogenic probe and standards. |
| pH Range / Optimal pH | Assay buffer pH 7.5; MPA deproteinization at acidic pH. |
| Shipping Conditions | Cold pack (enzyme and probe on dry ice recommended). |
| Expiration Date / Stability | 6 months at -20 °C; reconstituted probe stable 2 weeks at 2-8 °C. |
| Regulatory / Compliance | For research use only. |
| Compatibility | Compatible with cell and tissue lysates deproteinized with MPA. SDS and other detergents must be removed by deproteinization. Hemoglobin interferes with fluorescence — avoid hemolyzed blood samples. |
| Recommended Buffer System | Phosphate buffer with EDTA, pH 7.5. |
| Application Notes / Precautions | Deproteinization is critical — protein thiols interfere with probe and cause high background. Keep all samples on ice and process quickly as GSH auto-oxidizes to GSSG at neutral pH. Include GSH-only and GSSG-only controls for derivatization verification. Normalize results to protein content from parallel aliquot. For tissue, add 5-sulfosalicylic acid during homogenization to prevent GSH oxidation. |
| Batch-to-Batch Consistency | GSH standard concentration within ±5% by DTNB assay; probe reactivity within ±15% of reference lot. |
For research use only, not for clinical use.
|
There is no product in your cart. |
Copyright © 2026 Alta DiagnoTech. All rights reserved.
