Total Price: $0.00
Contact Information
Reduced Glutathione (GSH) Colorimetric Assay Kit
SKU: E-BC-K030-M-500
Reduced Glutathione (GSH) Colorimetric Assay Kit
| SKU # | E-BC-K030-M |
| Detection Instrument | Microplate reader(405-414 nm) |
| Detection Method | Colorimetric method |
Product Details
Properties
| Synonyms | GSH |
| Sample Type | Serum, plasma, cell culture supernatant, tissue, cells |
| Sensitivity | 2 μmol/L |
| Detection Range | 2-100 μmol/L |
| Detection Method | Colorimetric method |
| Assay type | Quantitative |
| Assay time | 25 min |
| Precision | Average inter-assay CV: 3.200% | Average intra-assay CV: 1.900% |
| 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 the pre-experiment and the detection range (2-100 μmol/L).
The recommended dilution factor for different samples is as follows (for reference only):
| Sample type | Dilution factor |
| Human serum | 1 |
| Human plasma | 1 |
| 10% Mouse brain tissue homogenization | 1 |
| 10% Mouse liver tissue homogenization | 1 |
| Hela cell homogenization（0.999 mgprot/mL） | 1 |
| Rat serum | 1 |
| Rat plasma | 1 |
| Mouse serum | 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 GSH can react with Dinitrobenzoic acid (DNTB) to form a yellow complex which can be detected by colorimetric assay at 405 nm and calculate the reduced GSH content indirectly.
Kit Components & Storage
| Item | Component | Size 1(48 T) | Size 2(96 T) | Storage |
| Reagent 1 | Acid Reagent | 6 mL × 1 vial | 12 mL × 1 vial | 2-8℃, 12 months shading light |
| Reagent 2 | Phosphate | 6 mL × 1 vial | 12 mL × 1 vial | 2-8℃, 12 months |
| Reagent 3 | DTNB Solution | 1.5 mL × 1 vial | 1.5 mL × 2 vials | 2-8℃, 12 months shading light |
| Reagent 4 | GSH Standard | 3.07 mg × 1 vial | 3.07 mg × 2 vials | 2-8℃, 12 months |
| Reagent 5 | GSH Standard Stock Diluent | 1.5 mL× 1 vial | 1.5 mL× 2 vials | 2-8℃, 12 months |
| Microplate | 96 wells | No requirement | ||
| Plate Sealer | 2 pieces | |||
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 (μmol/L) | 5.50 | 26.40 | 64.50 |
| %CV | 2.3 | 2.0 | 1.4 |
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 (μmol/L) | 5.50 | 26.40 | 64.50 |
| %CV | 3.5 | 2.9 | 3.2 |
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 96%.
| Standard 1 | Standard 2 | Standard 3 | |
| Expected Conc. (μmol/L) | 15 | 45 | 58 |
| Observed Conc. (μmol/L) | 14.9 | 42.3 | 55.1 |
| Recovery rate (%) | 99 | 94 | 95 |
Sensitivity
The analytical sensitivity of the assay is 2 μmol/L GSH 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.
Standard Curve:
As the OD value of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique or temperature effects), so the standard curve and data are provided as below for reference only:
| Concentration (μmol/L) | 0 | 10 | 20 | 40 | 50 | 60 | 80 | 100 |
| Average OD | 0.057 | 0.088 | 0.129 | 0.213 | 0.296 | 0.296 | 0.392 | 0.484 |
| Absoluted OD | 0.000 | 0.030 | 0.071 | 0.156 | 0.196 | 0.239 | 0.334 | 0.426 |