Reduced Glutathione (GSH) Colorimetric Assay Kit
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Reduced Glutathione (GSH) Colorimetric Assay Kit
| SKU # | E-BC-K030-S |
| Detection Instrument | Spectrophotometer(420 nm) |
| Detection Method | Colorimetric method |
Product Details
Properties
| Synonyms | GSH |
| Sample Type | Serum, plasma, tissue,cells |
| Sensitivity | 0.26 mg GSH/L |
| Detection Range | 0.26-122.8 mgGSH/L |
| Detection Method | Colorimetric method |
| Assay type | Quantitative |
| Assay time | 50 min |
| Precision | Average inter-assay CV: 2.400% | Average intra-assay CV: 1.800% |
| Other instruments required | Micropipettor, Vortex mixer |
| Other reagents required | Normal saline (0.9% NaCl) or PBS (0.01 M, pH 7.4) |
| Storage | 2-8℃ |
| Valid period | 12 months |
Dilution of Sample
It is recommended to take 2~3 samples with expected large difference to do pre-experiment before formal experiment and dilute the sample according to the result of pre-experiment and the detection range ( 0.26-122.8 mg GSH/L).
The recommended dilution factor for different samples is as follows (for reference only):
| Sample type | Dilution factor |
| Human serum | 1 |
| Mouse serum | 1 |
| 10% Mouse brain tissue homogenization | 1 |
| 10% Rat liver tissue homogenization | 1 |
| Human plasma | 1 |
| Rat plasma | 1 |
| 10% Carrot tissue homogenization | 1 |
| 293T supernatant | 1 |
Note: The diluent is normal saline (0.9% NaCl) or PBS (0.01 M, pH 7.4).
Detection Principle
Reduced glutathione (GSH) can react with dithionitrobenzoic acid (DTNB) to produce thio-nitrobenzoic acid and glutathione disulfide. Nitromercaptobenzoic acid is a yellow compound which has the maximum absorption peak at 420 nm. The GSH content can be calculated by measuring the OD value at 420 nm.
Kit Components & Storage
| Item | Component | Size 1 (50 assays) | Size 2 (100 assays) | Storage |
| Reagent 1 | Acid Reagent | 45 mL × 1 vial | 45 mL × 2 vials | 2-8℃, 12 months shading light |
| Reagent 2 | Phosphate | Powder × 1 vial | Powder × 2 vials | 2-8℃, 12 months |
| Reagent 3 | DTNB Solution | 1.5 mL × 1 vial | 30 mL × 1 vial | 2-8℃, 12 months shading light |
| Reagent 4 | Salt Reagent | Powder × 2 vials | Powder × 4 vials | 2-8℃, 12 months shading light |
| Reagent 5 | GSH Standard | 3.07 mg × 1 vial | 3.07 mg ×2 vials | 2-8℃, 12 months |
| Reagent 6 | GSH Standard Stock Diluent |
6 mL× 1 vial | 6 mL× 1 vial | 2-8℃, 12 months |
Note: The reagents must be stored strictly according to the preservation conditions in the above table. The reagents in different kits cannot be mixed with each other. For a small volume of reagents, please centrifuge before use, so as not to obtain sufficient amount of reagents.
Technical Data:
Parameter:
Intra-assay Precision
Three human serum samples were assayed in replicates of 20 to determine precision within an assay (CV = Coefficient of Variation).
| Parameters | Sample 1 | Sample 2 | Sample 3 |
| Mean (mg GSH/L) | 3.60 | 37.50 | 76.20 |
| %CV | 2.3 | 1.9 | 1.2 |
Inter-assay Precision
Three human serum samples were assayed 20 times in duplicate by three operators to determine precision between assays.
| Parameters | Sample 1 | Sample 2 | Sample 3 |
| Mean (mg GSH/L) | 3.60 | 37.50 | 76.20 |
| %CV | 2.8 | 2.0 | 2.4 |
Recovery
Take three samples of high concentration, middle concentration and low concentration to test the samples of each concentration for 6 times parallelly to get the average recovery rate of 102%.
| Sample 1 | Sample 2 | Sample 3 | |
| Expected Conc. (mg GSH/L) | 16 | 45.5 | 85.4 |
| Observed Conc. (mg GSH/L) | 16.5 | 45.5 | 88.0 |
| Recovery rate (%) | 103 | 100 | 103 |
Sensitivity
The analytical sensitivity of the assay is 0.26 mg GSH/L. This was determined by adding two standard deviations to the mean O.D. obtained when the zero standard was assayed 20 times, and calculating the corresponding concentration.