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Glucose-6-Phosphate (G6P) Colorimetric Assay Kit
SKU: E-BC-K011-M-96
Glucose-6-Phosphate (G6P) Colorimetric Assay Kit
| SKU # | E-BC-K011-M |
| Detection Instrument | Microplate reader (450 nm) |
| Detection Method | Colorimetric method |
Product Details
Properties
| Synonyms | G6P |
| Sample Type | Serum, plasma, animal tissue |
| Sensitivity | 5.6 μmol /L |
| Detection Range | 5.6-500 μmol/L |
| Detection Method | Colorimetric method |
| Assay type |
Quantitative |
| Assay Time | 35 min |
| Precision | Average inter-assay CV: 4.300% | 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 (5.6-500 μmol/L).
The recommended dilution factor for different samples is as follows (for reference only):
| Sample type | Dilution factor |
| Human plasma | 1 |
| Mouse serum | 02-Mar |
| Rat plasma | 1 |
| Porcine serum | 1 |
| 10% Rat spleen tissue homogenate | 02-Mar |
| 10% Rat heart tissue homogenate | 02-Mar |
| 10% Rat liver tissue homogenate | 02-Mar |
| 10% Mouse lung tissue homogenate | 1 |
Note: The diluent is reagent 1.
Detection Principle
Glucose-6-phosphate (G6P) is a molecule generated by phosphorylation of hydroxyl groups on the sixth carbon of glucose under the catalysis of hexokinase. It is a common small molecule of sugar metabolism in cells and participates in biochemical pathways such as glycolysis and pentose phosphate pathway. In the first reaction of glycolysis, glucose is catalyzed by hexokinase to produce glucose-6-phosphate, which is then catalyzed by phosphoglucose isomerase to form fructose-6-phosphate to continue the other steps of glycolysis: In the pentose phosphate pathway, glucose-6-phosphate is the first substrate, and this process is also the main way to generate NADPH. In addition to these two metabolic pathways, glucose-6-phosphate can also be converted into glycogen or starch and stored.
Kit Components & Storage
| Item | Component | Size (96 T) | Storage |
| Reagent 1 | Extracting Solution | 60 mL×2 vials | -20℃, 12 months |
| Reagent 2 | Buffer Solution | 5 mL × 1 vial | -20℃, 12 months |
| Reagent 3 | Chromogenic Agent | 1.5 mL × 2 vials | -20℃, 12 months, shading light |
| Reagent 4 | Enzyme Agent | Powder × 1 vial | -20℃, 12 month |
| Reagent 5 | 10 mmol/L G6P Standard | 0.5 mL × 1 vial | -20℃, 12 months |
| 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 (μmol/L) |
10.00 | 185.00 | 375.00 |
| %CV | 2.6 | 2.0 | 1.7 |
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 (μmol/L) |
10.00 | 185.00 | 375.00 |
| %CV | 4.2 | 4.5 | 4.2 |
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 95%.
| Standard 1 | Standard 2 | Standard 3 | |
| Expected Conc.(μmol/L) | 85 | 265 | 385 |
| Observed Conc.(μmol/L) | 83.3 | 251.8 | 354.2 |
| Recovery rate (%) | 98 | 95 | 92 |
Sensitivity
The analytical sensitivity of the assay is 5.6 μmol/L G6P. 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 | 200 | 300 | 350 | 400 | 500 |
| Average OD | 0.055 | 0.158 | 0.324 | 0.608 | 0.845 | 1.027 | 1.179 | 1.442 |
| Absoluted OD | 0.000 | 0.103 | 0.269 | 0.553 | 0.791 | 0.973 | 1.124 | 1.388 |