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Glucose-6-Phosphate Dehydrogenase (G-6-PD) Activity Assay Kit
SKU: E-BC-K056-M-96
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Glucose-6-Phosphate Dehydrogenase (G-6-PD) Activity Assay Kit
| SKU # | E-BC-K056-M |
| Detection Instrument | Microplate reader(450 nm) |
| Detection Method | Colorimetric method |
Product Details
Properties
| Synonyms | G-6-PD, G6PDH |
| Sample Type | Serum, plasma, animal tissue |
| Sensitivity | 0.01 U/L |
| Detection Range | 0.01-50 U/L |
| Detection Method | Colorimetric method |
| Assay type | Enzyme Activity |
| Assay time | 20 min |
| Precision | Average inter-assay CV: 5.800% | Average intra-assay CV: 2.100% |
| Other instruments required | Pipettor, Water bath, Centrifuge |
| Storage | -20℃ |
| Valid period | 12 months |
Dilution of Sample
It is recommended to take 2~3 samples with expected large difference to do pre-experiment before formal experiment and dilute the sample according to the result of the pre-experiment and the detection range (0.01-50 U/L).
The recommended dilution factor for different samples is as follows (for reference only):
| Sample type | Dilution factor |
| Human serum | 1 |
| Rat serum | 1 |
| Rabbit serum | 1 |
| Cynomolgus monkey | 1 |
| 10% Mouse liver tissue homogenate | 3-5 |
| 10% Rat kidney tissue homogenate | 1 |
| 10% Rat spleen tissue homogenate | 5-10 |
| 10% Mouse brain tissue homogenate | 1 |
Note: The diluent is reagent 1.
Detection Principle
Under the presence of G6PDH, glucose-6-phosphoric acid is oxidized to 6-PG. In this reaction, NADP+ is reduced to NADPH. Under the action of electron coupling reagent 1-MPMS, NADPH reducts wST-8 to form orange formazan, which has the maximum absorption peak at about 450 nm. Formazan generated in the reaction system is proportional to the activity of G6DPH in the sample.
Kit Components & Storage
| Item | Component | Size (96 T) | Storage |
| Reagent 1 | Extracting Solution | 50 mL ×2 vials | -20℃, 12 months |
| Reagent 2 | Substrate | 1.5 mL × 2 vials | -20℃, 12 months |
| Reagent 3 | Chromogenic Agent | 1.5 mL × 2 vials | -20℃, 12 months, shading light |
| Reagent 4 | Buffer Solution | 4 mL × 1 vial | -20℃, 12 months, shading light |
| Reagent 5 | 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) | 2.60 | 11.50 | 38.50 |
| %CV | 2.4 | 2.0 | 1.9 |
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) | 2.60 | 11.50 | 38.50 |
| %CV | 5.7 | 6.1 | 5.6 |
Sensitivity
The analytical sensitivity of the assay is 0.01 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 | 50 | 100 | 150 | 250 | 300 | 400 | 500 |
| Average OD | 0.059 | 0.182 | 0.323 | 0.447 | 0.724 | 1.019 | 1.190 | 1.463 |
| Absoluted OD | 0.000 | 0.124 | 0.265 | 0.388 | 0.666 | 0.961 | 1.132 | 1.404 |