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| Catalog # | Availability | Size | Quantity | Unit Price | Save For Later Wish List | |
|---|---|---|---|---|---|---|
| EKFT19768UV-48 | 2 weeks | 48T | $168.00 |
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| EKFT19768UV-96 | 2 weeks | 96T | $240.00 |
Product Overview | ||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Name | Cr (Creatinine) Assay Kit (Colorimetric) | |||||||||||||||||||||||||||||||||||||||||||||||||||||
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| Sensitivity | 10μmol/L | |||||||||||||||||||||||||||||||||||||||||||||||||||||
| Sample type(s) | Serum, Plasma, Cell Culture Supernatant, cell or tissue lysate, Other liquid samples | |||||||||||||||||||||||||||||||||||||||||||||||||||||
| Precision | ||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Intra-Assay: CV≤6% ,Inter-Assay: CV≤4% | ||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Recovery | ||||||||||||||||||||||||||||||||||||||||||||||||||||||
Matrices listed below were spiked with certain level of creatinine and the recovery rates were calculated by comparing the measured value to the expected amount of creatinine in samples.
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| Stability | ||||||||||||||||||||||||||||||||||||||||||||||||||||||
| The kit is stored at 2-8°C (Avoid Direct Light), and not be frozen or thawed. The product is valid for 6 months. After opening, store at 2-8°C, it can be stable for 30 days, avoid contamination. (human serum：60-120umol/L,)Operation Procedure Principle of the Assay Creatinine is catalyzed by creatininase to form creatine, and creatine is catalyzed by creatinase to form sarcosine and urea, then sarcosine catalyzed by sarcosine oxidase to form glycine, formaldehyde and hydrogen peroxide. Hydrogen peroxide and chromogenic agent react under the catalysis of peroxidase to produce pink compound The creatinine content was calculated by ready the OD value at 546 nm( )/700 nm (Deputy wavelength). Precautions To inspect the validity of experiment operation and the appropriateness of sample dilution proportion, pilot experiment using standards and a small number of samples is recommended. Before using the kit, spin tubes and bring down all components to the bottom of tubes. Storage all reagents avoid light. Duplicate well assay is recommended for both standard and sample testing. Don’t reuse tips and tubes to avoid cross contamination. Avoid using the reagents from different batches together. Material Required but Not Supplied Microplate reader (wavelength:546nm/700nm) 37°C incubator Vortex mixer Precision single and multi-channel pipette and disposable tips Clean tubes and Eppendorf tubes Deionized or distilled water. Sample Collection and Storage (universal) Serum: Place whole blood sample at room temperature for 2 hours or put it at 2-8°C overnight and centrifugation for 20 minutes at approximately 1000×g, Collect the supernatant and carry out the assay immediately. Blood collection tubes should be disposable, non-pyrogenic, and non-endotoxin. Plasma: Collect plasma using EDTA-Na2 or heparin as an anticoagulant. Centrifuge samples for 15 minutes at 1000×g at 2-8°C within 30 minutes of collection. Collect the supernatant and carry out the assay immediately. Avoid hemolysis, high cholesterol samples. Urine: Centrifuge supernatant for 20 minutes at 1000×g at 2-8°C to remove insoluble impurity and cell debris. Collect the clear supernatant and carry out the assay immediately. Note: Samples to be used within 5 days can be stored at 2-8°C, besides that, samples must be stored at -20°C (assay ≤1 month) or -80°C(assay≤2 months) to avoid loss of bioactivity and contamination. Sample Dilution The user should estimate the concentration of the creatinine in the test sample, and select a proper dilution factor to make the diluted target protein concentration fall in the optimal detection range of the kit. Dilute the sample with distilled water, and several trials may be necessary. The test sample must be well mixed with distilled water. And also standard curves and sample should be making in pre-experiment. Reagent Preparation and Storage Bring all reagents and samples to room temperature for 20 minutes before use. 1, Sample prepare:
2，Standard prepare: Diluted standard solution is unstable and must be used within 2 hours. 1, Add 1 ml distilled water into Standard tube (labeled as stock), keep the tube at room temperature for 10 minutes and mix them thoroughly. 2, Refer to the table to prepare the standard.
The following two assay procedures are provided, please choose by yourself, the results are all for reference. Assay Procedure(Standard operating procedure) Set standard, test samples control (blank) wells on the plate respectively, and then, records their positions. It is recommended to measure each standard and sample in duplicate. Prepare Standards: Aliquot 10μl of standard (1-7) and blank into the standard wells. Add Samples: Add 10μl of properly diluted sample into sample wells. Add Enzyme Solution A: Add 210μl of enzyme Solution A into each well and mix the well. Incubate: Seal the plate with a cover and incubate at 37°C for 5 minutes. OD Measurement: Remove the cover and read the O.D. absorbance at 546nm in Microplate Reader immediately(mark A1) Add Enzyme Solution B: Add 70μl of enzyme Solution B into each well and mix the well. Incubate: Seal the plate with a cover and incubate at 37°C for 5 minutes. OD Measurement: Remove the cover and read the O.D. absorbance at 546nm in Microplate Reader immediately(mark A2) Calculation: Use the software for calculation Regarding calculation, (the relative O.D.546) = (the O.D.546 A2 of each well) – (the O.D.546 A1 of each well)). The standard curve can be plotted as the relative O.D.546 of each standard solution (Y) vs. the respective concentration of the standard solution (X). The target concentration of the samples can be interpolated from the standard curve. It is recommended to use some professional software to do this calculation, such as Curve Expert 1.3 or 1.4. Assay Procedure(Simple operating procedure) 1. Set standard (Stock), test samples, control (blank) wells on the plate respectively, and then, records their positions. It is recommended to measure each standard and sample in duplicate. 2. Prepare Standard: Aliquot 10μl of Stock Standard and blank into the standard/blank wells. 3. Add Samples: Add 10μl of properly diluted sample into test sample wells. Add Enzyme Solution A: Add 210μl of enzyme Solution A into each test well mix the well. Incubate: Seal the plate with a cover and incubate at 37°C for 5 minutes. OD Measurement: Remove the cover and read the O.D. absorbance at 546nm in Microplate Reader immediately(A1) Add Enzyme Solution B: Add 70μl of enzyme Solution B into each test well mix the well. Incubate: Seal the plate with a cover and incubate at 37°C for 5 minutes. OD Measurement: Remove the cover and read the O.D. absorbance at 546nm in Microplate Reader immediately(A2) Calculation:  Note: If the samples measured were diluted, multiply the dilution factor to the concentrations from interpolation to obtain the concentration before dilution. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||
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