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Glycosylated Serum Protein (GSP) Colorimetric Assay Kit
SKU: E-BC-K069-M-96
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Glycosylated Serum Protein (GSP) Colorimetric Assay Kit
| SKU # | E-BC-K069-M |
| Detection Instrument | Microplate reader (520-550 nm, optimum wavelength: 530 nm) |
| Detection Method | Colorimetric method |
Product Details
Properties
| Synonyms | GSP |
| Sample Type | Serum, plasma |
| Sensitivity | 0.06 mmol/L |
| Detection Range | 0.06 - 4.0 mmol/L |
| Detection Method | Colorimetric method |
| Assay type | Quantitative |
| Assay time | 30 min |
| Precision | Average inter-assay CV: 6.300% | Average intra-assay CV: 4.400% |
| Other instruments required | Incubator |
| 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 |
| Human serum | 1 |
| Rat serum | 1 |
| Mouse serum | 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
GSP with ketoamine structure is able to react with tetrazole blue under alkaline conditions to form purplish red product, which has a characteristic absorption peak at 530 nm, the concentration of GSP can be calculated by measuring the OD value.
Kit Components & Storage
| Item | Component | Size 1(48 T) | Size 2(96 T) | Storage |
| Reagent 1 | Standard Diluent | 0.5 mL ×1 vial | 0.5 mL ×1 vial | -20℃, 12months |
| Reagent 2 | 2 mmol/L Standard Solution | 0.5 mL ×1 vial | 0.5 mL ×1 vial | -20℃, 12months |
| Reagent 3 | Chromogenic Agent | 12 mL ×1 vial | 25 mL × 1 vial | -20℃, 12 months, shading light |
| Reagent 4 | Stop Solution | 3 mL ×1 vial | 6 mL × 1 vial | -20℃, 12months |
| 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 (mmol/L) | 0.85 | 2.30 | 3.40 |
| %CV | 4.6 | 4.0 | 4.6 |
Inter-assay Precision
Three human serum samples were assayed 17 times in duplicate by three operators to determine precision between assays.
| Parameters | Sample 1 | Sample 2 | Sample 3 |
| Mean (mmol/L) | 0.85 | 2.30 | 3.40 |
| %CV | 6.5 | 6.1 | 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. (mmol/L) | 1.2 | 2.7 | 3.6 |
| Observed Conc. (mmol/L) | 1.2 | 2.8 | 3.8 |
| Recovery rate (%) | 102 | 105 | 105 |
Sensitivity
The analytical sensitivity of the assay is 0.06 mmol/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 | 0.5 | 1 | 1.5 | 2 | 3 | 4 | 5 |
| OD value | 0.076 | 0.099 | 0.124 | 0.152 | 0.170 | 0.224 | 0.280 | 0.339 |
| 0.076 | 0.104 | 0.121 | 0.146 | 0.175 | 0.210 | 0.267 | 0.335 | |
| Average OD | 0.076 | 0.102 | 0.123 | 0.149 | 0.173 | 0.217 | 0.274 | 0.337 |
| Absoluted OD | 0.000 | 0.026 | 0.047 | 0.073 | 0.097 | 0.141 | 0.198 | 0.261 |