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β-N-Acetylglucosaminidase (NAG) Activity Assay Kit
SKU: E-BC-K064-M-96
β-N-Acetylglucosaminidase (NAG) Activity Assay Kit
| SKU # | E-BC-K064-M |
| Detection Instrument | Microplate reader (390-415 nm, optimum wavelength: 400 nm) |
| Detection Method | Colorimetric method |
Product Details
Properties
| Synonyms | NAG |
| Sample Type | Serum, plasma, animal tissue and cell |
| Sensitivity | 0.76 U/L |
| Detection Range | 0.76 - 49.51 U/L |
| Detection Method | Colorimetric method |
| Assay type | Enzyme Activity |
| Precision | Average inter-assay CV: 4.6-6.8% | Average intra-assay CV: 3.2-5.0% |
| Other instruments required | Incubator(37℃) |
| Other reagents required | Double distilled water, Normal saline(0.9% NaCl) |
| 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 | 3-5 |
| 10% Mouse kidney tissue homogenate | 3-5 |
| 10% Mouse heart tissue homogenate | 3-5 |
| 10% Mouse lung tissue homogenate | 3-5 |
| Rat plasma | 1 |
| Human serum | 1 |
| Bovine serum | 2-3 |
| 6×10^6 CHO cells | 1 |
| 0.6×10^6 293T/17 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
β-N-Acetylglucosaminidase (NAG) catalyzed the substrate to produce p-nitrophenol, which has an absorption wavelength at 400 nm. Therefore, the activity of NAG can be calculated by measuring the change of absorbance value at 400 nm.
Kit Components & Storage
| Item | Component | Size (96 T) | Storage |
| Reagent 1 | Buffer Solution | 20 mL ×1 vial | -20℃, 12 months |
| Reagent 2 | Substrate Solution | 5 mL × 1 vial | -20℃, 12 months, shading light |
| Reagent 3 | Chromogenic Agent | 14 mL × 1 vial | -20℃, 12 months |
| Reagent 4 | Standard | Powder × 1 vial | -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) | 0.50 | 2.50 | 5.00 |
| %CV | 5.0 | 3.2 | 4.4 |
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 (U/L) | 0.50 | 2.50 | 5.00 |
| %CV | 6.8 | 4.6 | 4.8 |
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. (mmol/L) | 0.12 | 0.25 | 0.38 |
| Observed Conc. (mmol/L) | 0.1 | 0.2 | 0.4 |
| Recovery rate (%) | 92 | 98 | 98 |
Sensitivity
The analytical sensitivity of the assay is 0.76 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 (mmol/L) | 0 | 0.1 | 0.15 | 0.2 | 0.3 | 0.35 | 0.4 | 0.5 |
| OD Value | 0.064 | 0.161 | 0.218 | 0.267 | 0.372 | 0.435 | 0.471 | 0.573 |
| 0.065 | 0.161 | 0.218 | 0.269 | 0.374 | 0.427 | 0.478 | 0.572 | |
| Average OD | 0.065 | 0.161 | 0.218 | 0.268 | 0.373 | 0.431 | 0.475 | 0.573 |
| Absoluted OD | 0.000 | 0.097 | 0.154 | 0.204 | 0.309 | 0.367 | 0.410 | 0.508 |