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Creatinine (Cr) Colorimetric Assay Kit (Sarcosine Oxidase Method)
SKU: E-BC-K188-M-500
Creatinine (Cr) Colorimetric Assay Kit (Sarcosine Oxidase Method)
| SKU # | E-BC-K188-M |
| Detection Instrument | Microplate reader (510-520 nm, optimum wavelength: 515 nm) |
| Detection method | Colorimetric method |
Product Details
Properties
| Synonyms | Cr |
| Sample Type | Serum, plasma, urine |
| Sensitivity | 3.8 μmol/L |
| Detection Range | 20.45-400 μmol/L |
| Detection Method | Colorimetric method |
| Assay type | Quantitative |
| Assay time | 30 min |
| Precision | Average inter-assay CV: 3.700% | Average intra-assay CV: 1.400% |
| Other instruments required | Micropipettor, Centrifuge, Incubator, Vortex mixer |
| Other reagents required | Normal saline (0.9% NaCl) |
| Storage | 2-8℃ |
| Valid period | 12 months |
Images
X Zhang et al investigate the usage of H2O2 and O2 for cooperative chemodynamic / starvation cancer therapy. Creatinine (Cr) level in mouse blood was determined using creatinine colorimetric assay kit (E-BC-K188-M).
No significant change of creatinine was observed, the LCFG nanoparticles did not include nephrotoxicity.
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 (38.2-800 μmol/L).
The recommended dilution factor for different samples is as follows (for reference only):
| Sample type | Dilution factor |
| Human urine | 20-30 |
| Human serum | 1 |
| Rat serum | 1 |
| Porcine serum | 1 |
Note: The diluent is normal saline (0.9% NaCl).
Detection Principle
Creatinine (Cr) can be catalyzed by creatinase and generates creatine. Creatine can be hydrolyzed into sarcosine and urea by creatinase. The sarcosine can be catalyzed by sarcosine oxidase and form glycine, formaldehyde and hydrogen peroxide. The reaction between hydrogen peroxide, 2,4-(6-Tri-iodine-3- hydroxybenzoic acid) and 4-ampyrone can be catalyzed by peroxidase and form pink compound. Creatinine content can be calculated indirectly by measuring the OD value at 515 nm.
Kit Components & Storage
| Item | Component | Size 1(48 T) | Size 2(96 T) | Storage |
| Reagent 1 | Enzyme Solution A | 10 mL × 1 vial | 20 mL × 1 vial | 2-8°C, 12 months, shading light |
| Reagent 2 | Enzyme Solution B | 3.5 mL × 1 vial | 7 mL × 1 vial | 2-8°C, 12 months, shading light |
| Reagent 3 | 1 mmol/L Standard Solution | 1.5 mL × 1 vial | 1.5 mL × 2 vials | 2-8°C, 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) | 88.60 | 271.00 | 563.00 |
| %CV | 1.7 | 1.5 | 1.0 |
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) | 88.60 | 271.00 | 563.00 |
| %CV | 3.2 | 3.7 | 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 106%.
| Standard 1 | Standard 2 | Standard 3 | |
| Expected Conc. (mmol/L) | 0.13 | 0.22 | 0.34 |
| Observed Conc. (mmol/L) | 0.1 | 0.2 | 0.4 |
| Recovery rate (%) | 108 | 106 | 104 |
Sensitivity
The analytical sensitivity of the assay is 3.8 μmol/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 | 0.25 | 0.3 | 0.35 | 0.4 |
| Average ΔA | 0.001 | 0.042 | 0.057 | 0.078 | 0.097 | 0.124 | 0.144 | 0.171 |
| Absoluted OD | 0.000 | 0.042 | 0.056 | 0.078 | 0.096 | 0.123 | 0.143 | 0.171 |