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Cat.No: DBX-QCY-1681 Datasheet Instruction for Use
Specification Quantities
Price $752-1860
| Product Name | Human Ferritin ELISA Kit |
| Catalog No. | DBX-QCY-1681 |
| Description | The kit employs an enzyme-linked immunosorbent assay (ELISA) based on a double-antibody sandwich method. It determines the concentration of human ferritin in a sample by measuring the absorbance of the sample in the reaction wells, based on the principle that ferritin binds specifically to the coated antibodies on the microplate. |
| Sample Type | Serum, plasma, cell culture supernatant |
| Sensitivity | 46.88 pg/mL |
| Detection Range | 93.75–6000 pg/mL |
| Assay Method | Sandwich ELISA |
| Reagent Preparation | Reagent Warm-up: Place the test kit and samples to be tested at room temperature 30 minutes before the experiment. If crystals form in the concentrated wash solution, place it in a 37°C water bath until all crystals have dissolved. Preparation of Wash Buffer: Calculate the required volume of diluted wash buffer in advance, then dilute the 20× concentrated wash buffer with double-distilled water ordeionized water to prepare the 1× working solution. Store any unused concentrated wash bufferin a 4°C refrigerator. Gradient Dilution of Standards: Before opening the container, ensure that all lyophilized standards are settled at the bottom. Add 0.5 mL of SR1 standard/sample diluent to the lyophilized standards, let stand for 10–30 minutes until completely dissolved, then gently mix. Mix thoroughly before dilution, and perform a 2-fold dilution according to the following concentrations: 6000, 3000, 1500, 750, 375, 187.5, and 93.75 pg/mL. 6000 pg/mL is the highest concentration on the standard curve, and the SR1 standard/sample diluent serves as the blank (0 pg/mL) on the standard curve. Any unused reconstituted stock solution (6000 pg/mL) should be discarded or aliquoted into single-use portions as needed, and stored in a freezer at -20°C to -80°C. Biotinylated Antibody Working Solution: Calculate the required volume for the current experiment in advance. Use SR2 biotinylated antibody diluent to dilute the 100× antibody concentrate to a 1× working solution (mix thoroughly before dilution). Add to the reaction wells within 30 minutes. Enzyme-conjugate working solution: Calculate the required volume for the current experiment in advance. Dilute the 40× enzyme-conjugate concentrate with SR3 enzyme-conjugate diluent to prepare a 1× working solution (mix thoroughly before dilution). Use within 30 minutes. Washing Procedure: Automated Washing: Centrifuge to remove liquid from the microplate wells, tap dry on thick absorbent paper, and dispense 300 μL of wash solution per well. Allow 30 seconds between dispensing and aspiration. Repeat this washing step according to the number of washes specified in the detection procedure. Manual washing: Centrifuge to remove liquid from the microplate wells, blot dry on thick absorbent paper, add 300 μL of wash solution per well using a wash bottle, let stand for 30 seconds, then centrifuge to remove liquid from the wells, blot dry on thick absorbent paper, and repeat this washing step according to the number of washes specified in the assay procedure. |
| Test Procedure | Before starting the assay, allow all reagents and samples to reach room temperature. Prepare all necessary reagents and working-concentration standards. Designate the standard wells, blank wells, and sample wells. Calculate the number of strips required for the current assay, remove the unused strips, reseal the package with a new sealing film, and return it to the aluminum foil pouch containing the desiccant. Soak the microplate: Add 300 μL of 1× wash buffer and let it soak for 30 seconds. Discard the wash buffer, then tap the microplate on absorbent paper to dry it. Repeat this process twice. Add standards: Add 100 μL of a 2-fold serial dilution of the standard to the standard wells. Add 100 μL of standard/sample diluent to the blank wells. Add samples: Add 100 μL of the sample to be tested to the sample wells. Ensure that steps 4 and 5 are performed consecutively without interruption. Complete the sample addition within 10 minutes. Incubation: Cover the plate with a new sealing film. Incubate at 37°C for 90 minutes. Wash: Discard the liquid. Add 300 μL of wash buffer to each well and wash the plate four times. After each wash, blot the wells dry on absorbent paper. Add biotinylated detection antibody: Add 100 μL of biotinylated antibody working solution to the reaction wells. Incubation: Cover the plate with a new sealing film. Incubate at 37°C for 60 minutes. Wash: Discard the liquid. Add 300 μL of wash buffer to each well and wash the plate four times. After each wash, blot dry on absorbent paper. Add enzyme conjugate: Add 100 μL of enzyme conjugate working solution to each reaction well. Incubation: Cover the plate with a new sealing film. After sealing, incubate at 37°C for 30 minutes. Wash: Discard the liquid. Add 300 μL of wash solution to each well and wash the plate 5 times. After each wash, blot dry on absorbent paper. Add Substrate for Color Development: Add 100 μL of TMB color development substrate to each well. Protect from light and incubate at 37°C in the dark for 15 minutes. Add stop solution: Add 50 μL of stop solution to each well. The order of adding the stop solution should, as much as possible, match the order in which the color development substrate was added. Measurement: Within 5 minutes, use a microplate reader to perform dual-wavelength detection, measuring the OD values at the maximum absorption wavelength of 450 nm and the reference wavelength of 630 nm. |
| Storage and Transportation | Store at 2–8°C for 6 months. Ship refrigerated. |
For research use only, not for clinical use.
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