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Sialic Acid (SA) Colorimetric Assay Kit
SKU: E-BC-K068-S-100
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Sialic Acid (SA) Colorimetric Assay Kit
| SKU # | E-BC-K068-S |
| Detection Instrument | Spectrophotometer (560 nm) |
| Detection Method | Colorimetric method |
Product Details
Properties
| Synonyms | SA |
| Sample Type | Serum, plasma, saliva, urine, hydrothorax, tissue, cells |
| Sensitivity | 0.022 mmol/L |
| Detection Range | 0.022-7 mmol/L |
| Detection Method | Colorimetric method |
| Assay type | Quantitative |
| Assay time | 60 min |
| Precision | Average inter-assay CV: 7.100% | Average intra-assay CV: 3.900% |
| Other instruments required | Micropipettor, Incubator, Vortex mixer, Centrifuge |
| Other reagents required | Normal saline (0.9% NaCl), PBS (0.01 M, pH 7.4) |
| Storage | Reagent 1: -20℃, Reagent 2: 2-8℃ |
| 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 (0.022-7 mmol/L).
The recommended dilution factor for different samples is as follows (for reference only):
| Sample type | Dilution factor |
| Human serum | 1 |
| Rat plasma | 1 |
| 10% Carrot tissue homogenization | 1 |
| Human saliva | 1 |
| 10% Mouse liver tissue homogenization | 1 |
| 10% Mouse brain tissue homogenization | 1 |
| Human hydrothorax | 1 |
Note: The diluent is normal saline (0.9% NaCl) or PBS (0.01 M, pH 7.4).
Detection Principle
Sialic acid forms a purplish red complex with methyl resorcinol in the presence of oxidant. And the absorbance conforms to Lambert-Beer's law. The content of sialic acid can be calculated by measuring the OD value at 560 nm. When detect tissue and cell samples, the protein concentration of the sample needs to be detected in addition (E-BC-K165, E-BC-K168, E-BC-K318 are recommended).
Kit Components & Storage
| Item | Component | Size 1 (50 assays) | Size 2 (100 assays) | Storage |
| Reagent 1 | 1 mmol/L SA Standard | 0.5 mL × 1 vial | 1 mL × 1 vial | -20℃, 12 months, shading light |
| Reagent 2 | Chromogenic Agent | 60 mL × 4 vials | 60 mL × 8 vials | 2-8℃, 12 months, shading light |
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 | 3.40 | 6.30 |
| %CV | 4.3 | 4.0 | 3.4 |
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 (mmol/L) | 0.85 | 3.40 | 6.30 |
| %CV | 6.9 | 7.0 | 7.4 |
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%.
| Sample 1 | Sample 2 | Sample 3 | |
| Expected Conc. (mmol/L) | 8 | 18 | 20 |
| Observed Conc. (mmol/L) | 7.4 | 17.6 | 19.0 |
| Recovery rate (%) | 92 | 98 | 95 |
Sensitivity
The analytical sensitivity of the assay is 0.022 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 | 1.0 | 2.0 | 3.0 | 5.0 | 6.0 | 7.0 |
| Average OD | 0.001 | 0.081 | 0.170 | 0.266 | 0.460 | 0.547 | 0.637 |
| Absoluted OD | 0 | 0.080 | 0.169 | 0.265 | 0.459 | 0.546 | 0.636 |