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Thioredoxin Reductase (TrxR) Activity Assay Kit
SKU: E-BC-K548-M-96
Thioredoxin Reductase (TrxR) Activity Assay Kit
| SKU # | E-BC-K548-M |
| Detection Instrument | Microplate reader (412 nm) |
| Detection method | Colorimetric method |
Product Details
Properties
| Synonyms | TrxR |
| Sample type | animal tissue and cell |
| Sensitivity | 0.82 U/L |
| Detection range | 0.82-46.81 U/L |
| Detection Method | Colorimetric method |
| Assay type | Enzyme Activity |
| Precision | Average inter-assay CV: 3.2-5.3% | Average intra-assay CV: 1.5-2.9% |
| Other instruments required | Incubator(37℃) |
| Storage | -20℃ |
| Valid period | 12 months |
Dilution of Sample
The recommended dilution factor for different samples is as follows (for reference only):
| Sample type | Dilution factor |
| 10% Mouse liver tissue homogenate | 2-3 |
| 10% Mouse kidney tissue homogenate | 2-3 |
| 10% Mouse lung tissue homogenate | 2-3 |
| HL-60 cells | 1 |
| CHO cells | 1 |
| Hela cells | 1 |
| Jurkat cells | 1 |
| Molt-4 cells | 1 |
Note: The diluent is normal saline (0.9% NaCl). For the dilution of other sample types, please do pretest to confirm the dilution factor.
Detection Principle
Thioredoxin system, as one of the most important antioxidant systems in the body, can stabilize the redox balance of organisms, regulate signal transduction. It is also related to cell REDOX reaction, nucleic acid metabolism, cell growth and tumorigenesis. Thioredoxin reductase (TrxR) is a NADPH-dependent dimer selenase containing the flavin adenine dinucleotide (FAD) domain and is a member of the pyridine nucleotide-disulfide oxidoreductase family. The thioredoxin system consists of NADPH, thioredoxin reductase and thioredoxin. The TrxR catalytic substrate makes the reducing agent reduce the color developer with a characteristic absorption peak at 412 nm.
Kit Components & Storage
| Item | Component | Size 1(48 T) | Size 2(96 T) | Storage |
| Reagent 1 | Buffer Solution | 25 mL × 1 vial | 50 mL × 1 vial | -20℃, 12 months |
| Reagent 2 | Substrate A | Powder × 1 vial | Powder × 2 vials | -20℃, 12 months shading light |
| Reagent 3 | Substrate B | Powder × 1 vial | Powder × 2 vials | -20℃, 12 months shading light |
| Reagent 4 | Chromogenic Agent | 0.25 mL × 1 vial | 0.25 mL × 2 vials | -20℃, 12 months shading light |
| 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 (U/L) | 2.4 | 18.6 | 32.5 |
| %CV | 2.9 | 2.8 | 1.5 |
Inter-assay Precision
Three human serum samples were assayed 17 times in duplicate by three operators to determine precision between assays.
| Parameters | Sample 1 | Sample 2 | Sample 3 |
| Mean (U/L) | 2.4 | 18.6 | 32.5 |
| %CV | 3.2 | 5.3 | 4.1 |
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%.
| Sample 1 | Sample 2 | Sample 3 | |
| Expected Conc. (μmol/L) | 10.5 | 22.5 | 38.3 |
| Observed Conc. (μmol/L) | 10.3 | 21.4 | 36.4 |
| Recovery rate (%) | 98 | 95 | 95 |
Sensitivity
The analytical sensitivity of the assay is 0.82 U/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.