Glutamate Synthase (GOGAT) Activity Assay Kit
Glutamate Synthase (GOGAT) Activity Assay Kit
| SKU # | E-BC-K659-M |
| Detection Instrument | Microplate reader (330-350 nm, optimum wavelength: 340 nm) |
| Detection method | Colorimetric method |
Product Details
Properties
| Synonyms | GOGAT |
| Sample type | Serum, plant and animal tissue |
| Sensitivity | 8.84 U/L |
| Detection range | 8.84–321.05 U/L |
| Detection Method | Colorimetric method |
| Assay type | Enzyme Activity |
| Assay time |
15 min |
| Precision | Average inter-assay CV: 5.800% | Average intra-assay CV: 2.500% |
| Other instruments required | Centrifuge |
| 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% Rat kidney tissue homogenate | 2-4 |
| 10% Rat liver tissue homogenate | 2-4 |
| 10% Rat heart tissue homogenate | 2-4 |
| 10% Mouse liver tissue homogenate | 2-4 |
| Bovine serum | 1 |
| 10% pleurotus cornucopiae tissue homogenate | 1 |
| 10% beech mushroom tissue homogenate | 1 |
Note: The diluent is extraction solution. For the dilution of other sample types, please do pretest to confirm the dilution factor.
Detection Principle
GOGAT catalysis the reaction that transfer the amino from glutamine to a-KG to form two molecules of glutamic acid using NADH as the electron donor. The decreasing rate of NADH that can be measured at 340 nm can reflect the activity of GOGAT.
Kit Components & Storage
| Item | Component | Size 1(48 T) | Size 2(96 T) | Storage |
| Reagent 1 | Extraction Solution | 50 mL × 1 vial | 50 mL × 2 vials | -20℃, 12 months |
| Reagent 2 | Buffer Solution | 16 mL × 1 vial | 26 mL × 1 vial | -20℃, 12 months |
| Reagent 3 | Substrate A | Powder × 2 vials | Powder × 2 vials | -20℃, 12 months, shading light |
| Reagent 4 | Substrate B | Powder × 2 vials | Powder × 2 vials | -20℃, 12 months, shading light |
| Reagent 5 | Chromogenic Agent | Powder × 2 vials | Powder × 2 vials | -20℃, 12 months, shading light |
| UV 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) | 13.50 | 84.50 | 167.00 |
| %CV | 2.9 | 2.4 | 2.2 |
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 (U/L) | 13.50 | 84.50 | 167.00 |
| %CV | 5.7 | 5.4 | 6.3 |
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 104%.
| Sample 1 | Sample 2 | Sample 3 | |
| Expected Conc. (U/L) | 56.7 | 148 | 252 |
| Observed Conc. (U/L) | 56.1 | 155.4 | 272.2 |
| Recovery rate (%) | 99 | 105 | 108 |
Sensitivity
The analytical sensitivity of the assay is 8.84 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..