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Glutaminase (GLS) Activity Assay Kit
SKU: E-BC-K660-M-96
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Glutaminase (GLS) Activity Assay Kit
| SKU # | E-BC-K660-M |
| Detection Instrument | Microplate reader (440-460 nm, optimum wavelength: 450 nm) |
| Detection method | Colorimetric method |
Product Details
Properties
| Synonyms | GLS |
| Sample type | Serum (plasma), animal, plant tissue |
| Sensitivity | 0.003 U/L |
| Detection range | 0.003-18.0 U/L |
| Detection method | Colorimetric method |
| Assay type | Enzyme Activity |
| Assay time | 80 min |
| Precision | Average inter-assay CV: 8.300% | Average intra-assay CV: 5% |
| Other instruments required | Test tube, Incubator, Centrifuge |
| Other reagents required | PBS (0.01 mol/L, pH 7.4) |
| Storage | |
| 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 | 1 |
| 10% Rat brain tissue homogenate | 1 |
| 10% Mouse kidney tissue homogenate | 1 |
| 10% Rat liver tissue homogenate | 1 |
| 10% Rat heart tissue homogenate 1 |
1 |
| 10% Rat kidney tissue homogenate | 1 |
| Human serum | 1 |
| Human plasma | 1-3 |
Note: The diluent is PBS (0.01 M, pH 7.4). For the dilution of other sample types, please do pretest to confirm the dilution factor.
Detection Principle
Glutamine is decomposed to produce glutamic acid under the action of glutaminase. Glutamic acid is further transformed by glutamic acid dehydrogenase. Meanwhile, NAD+ is reduced to NADH, Which under the action of hydrogen transmitter, transfer electrons to WST-8 to produce the yellow product. The activity of glutaminase can be calculated by measuring the change of absorbance value at 450 nm.
Kit Components & Storage
| Item | Component | Size (96 T) | Storage |
| Reagent 1 | Substrate A | Powder × 2 vials | -20℃, 12 months |
| Reagent 2 | Standard | Powder × 2 vials | -20℃, 12 months, shading light |
| Reagent 3 | Diluent | 4 mL×1 vial | -20℃, 12 months |
| Reagent 4 | Enzyme Reagent | Powder × 2 vials | -20℃, 12 months, shading light |
| Reagent 5 | Buffer Solution | 20 mL×1 vial | -20℃, 12 months |
| Reagent 6 | Substrate B | Powder × 2 vials | -20℃, 12 months, shading light |
| Reagent 7 | Accelerator | Powder × 1 vial | -20℃, 12 months, shading light |
| Reagent 8 | Accelerator | 1.5 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) | 0.26 | 2.80 | 13.50 |
| %CV | 5.4 | 5.2 | 4.7 |
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) | 0.26 | 2.80 | 13.50 |
| %CV | 8.2 | 7.9 | 8.8 |
Sensitivity
The analytical sensitivity of the assay is 0.003 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.
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 | 0.1 | 0.15 | 0.2 | 0.3 | 0.4 | 0.45 | 0.5 |
| Average OD | 0.067 | 0.175 | 0.224 | 0.298 | 0.423 | 0.535 | 0.577 | 0.625 |
| Absoluted OD | 0.000 | 0.108 | 0.158 | 0.231 | 0.357 | 0.468 | 0.511 | 0.559 |