Glutamate Synthase (GOGAT) Activity Assay Kit
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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 |
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Note: The diluent is extraction solution. For the dilution of other sample types, please do pretest to confirm the dilution factor.
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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.
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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 | |||
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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.
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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 |
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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..