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Cat.No: CS-DB-0031 Datasheet Instruction for Use
Specification Quantities
Price $1358.82–2264.70
| Product Name | Human CRP (C-Reactive Protein) ELISA Kit |
| Catalog No. | CS-DB-0031 |
| Description | This kit employs a double-antibody sandwich ELISA method. Anti-human CRP antibodies are used to coat the microplate. During the assay, samples (or standards) and horseradish peroxidase (HRP)-labeled anti-human CRP detection antibodies are added. The human CRP in the samples (or standards) binds to the coated antibodies and the HRP-labeled anti-human CRP detection antibodies, forming a sandwich-type immune complex. Unbound components are washed away. A color development substrate (TMB) is added; under the catalysis of horseradish peroxidase, TMB turns blue, and upon addition of the stop solution, it turns yellow. The OD value is measured at a wavelength of 450 nm using a microplate reader. The CRP concentration is directly proportional to the OD450 value, and the CRP concentration in the sample is calculated by plotting a standard curve. |
| Kit Components | Micro ELISA Plate: 98 wells × 12 strips Reference Standard: 2 vials Concentrated HRP Conjugate (100×): 1 vial, 60 μL Reference Standard & Sample Diluent: 3 vials, 20 mL HRP Conjugate Diluent: 1 vial, 14 mL Concentrated Wash Buffer (25×): 1 vial, 30 mL Substrate Reagent: 1 vial, 10 mL Stop Solution: 1 vial, 10 mL Plate Sealer: 5 sheets Manual: 1 set Certificate of Analysis: 1 set |
| Detection Limit | 52.1 pg/mL |
| Detection Range | 125 - 8000 pg/mL |
| Assay Method | ELISA |
| Sample Preparation | Serum: After allowing whole blood samples to stand at room temperature for 2 hours or at 4°C overnight, centrifuge at 1000×g for 20 minutes and use the supernatant for testing. Blood collection tubes must be single-use, endotoxin-free tubes. Plasma: EDTA-Na₂ is the recommended anticoagulant. Centrifuge at 1000×g for 15 minutes within 30 minutes of sample collection, then use the supernatant for testing. Avoid using samples with hemolysis or high lipid levels. |
| Test Procedure | Set up standard wells, blank wells, and sample wells. Add 50 μL of a serial dilution of the standard to the standard wells; add 50 μL of the standard and sample diluent to the blank wells; and add 50 μL of the sample to the remaining wells (it is recommended to set up duplicate wells for all samples and standards during the assay). Immediately add 50 μL of HRP-conjugated enzyme working solution to each well. Cover the microplate with a seal and incubate at 37°C for 60 minutes. Note: When dispensing samples, place them at the bottom of the well, avoiding contact with the well walls. Gently swirl to mix and prevent the formation of bubbles. The entire dispensing process should be completed within 10 minutes. Tap the plate to remove excess liquid from the wells, then blot dry on clean absorbent paper. Add 350 μL of wash buffer to each well, soak for 1 minute, then aspirate or tap out the liquid from the microplate and blot dry. Repeat this washing step 5 times. After washing is complete, proceed immediately to the next step; do not allow the microplate to dry out. Add 90 μL of substrate solution (TMB) to each well, cover the microplate with a seal, and incubate at 37°C in the dark for approximately 15 minutes. Note: Adjust the incubation time based on the actual color development, but do not exceed 30 minutes. Stop the reaction when a distinct gradient appears in the standard wells (a clear blue gradient appears in the first 4 wells). Turn on the microplate reader 15 minutes in advance to preheat it. Add 50 μL of stopping solution to each well to stop the reaction. Note: The order of adding the stopping solution should, as much as possible, match the order in which the substrate solution was added. Immediately measure the optical density (OD) of each well at a wavelength of 450 nm using the microplate reader. |
| Results | Calculate the mean OD values of the duplicate standard and sample wells, and subtract the OD value of the blank well to obtain the corrected value. Plot concentration on the x-axis and OD value on the y-axis, and fit a four-parameter logistic function to the standard curve on a double-logarithmic coordinate system. If a sample’s OD value exceeds the upper limit of the standard curve, the sample should be appropriately diluted and retested; when calculating the sample concentration, multiply the result by the corresponding dilution factor. |
| Applications | This kit is intended for the quantitative in vitro detection of CRP concentrations in human serum and plasma. |
| Storage and Transportation | Storage: 2-8°C. |
| Precautions | This kit is intended for in vitro research use only and is not intended for clinical diagnosis. Please wear a lab coat and latex gloves to ensure proper protection during the experiment. Newly opened microplate wells may contain a small amount of watery substance; this is normal and will not affect the experimental results. Strips not in immediate use should be removed from the plate and placed in the provided aluminum foil pouch, then stored according to the conditions listed in the table above. Do not reuse diluted standards or HRP-conjugated working solutions. Store unused concentrated HRP-conjugated solution (100×), microplates, and other undiluted reagents according to the storage conditions listed in the table above. The microplate reader used for testing must be equipped with a filter capable of detecting a wavelength of 450 ± 2 nm, with an optical density range of 0–3.5. It is recommended to preheat the instrument 15 minutes prior to use. Do not mix or substitute reagents from this kit with reagents from other lot numbers or sources. The Eppendorf tubes and tips used in the assay are for single-use only; mixing their use is strictly prohibited. Do not use expired reagents. Blood collection tubes must be single-use, endotoxin-free tubes. Avoid using samples with hemolysis or high lipid levels. If testing is performed within one week of sample collection, samples may be stored at 2–8°C. If testing cannot be performed promptly, aliquot samples into single-use portions and store at -20°C (for testing within one month) or -80°C (for testing within three months), avoiding repeated freeze-thaw cycles. Before testing, frozen samples should be thawed slowly and centrifuged to remove any precipitates formed during the freeze-thaw process. Mix thoroughly at room temperature before use. The assay range of the kit does not correspond to the concentration range of the analyte in the sample. If the analyte concentration in the sample is too high or too low, please dilute or concentrate the sample appropriately. |
For research use only, not for clinical use.
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