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Description
Human CYP24A1 ELISA KitProduct Specification Usage Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High precision pipette and pipette tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37 constant temperature box 4. Distilled water or deionized water Sample processing and requirements: Tissue, homogenates, cell lysates and other biological fluids Preparation before testing: 1. Please take out the test kit from the refrigerator 10 minutes in
Product Specification
| Usage |
Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High-precision pipette and pipette tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37℃ constant temperature box 4. Distilled water or deionized water Sample processing and requirements: Tissue, homogenates, cell lysates and other biological fluids Preparation before testing: 1. Please take out the test kit from the refrigerator 10 minutes in advance and equilibrate to room temperature. 2. Prepare a gradient standard working solution: Add 1 mL of universal diluent to the lyophilized standard. Let stand for 15 minutes to completely dissolve, then gently mix (concentration 10 ng/mL). Then dilute to the following concentrations: 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 ng/mL, 0.15625 ng/mL, and 0 ng/mL. Serial dilution method: Take seven EP tubes and add 500 μL of universal diluent to each tube. Pipette 500 μL of the 10 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 5 ng/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube serves as a blank well; there is no need to pipette liquid from the penultimate tube. See the figure below for details. 3. Preparation of biotinylated detection antibody working solution: Centrifuge the concentrated biotinylated antibody at 1000×g for 1 minute 15 minutes before use. Dilute the 100× concentrated biotinylated antibody to a 1× working concentration with universal diluent (e.g., 10uL concentrate + 990uL universal diluent). Prepare and use immediately. 4. Prepare the enzyme conjugate working solution: 15 minutes before use, centrifuge the 100× concentrated enzyme conjugate at 1000×g for 1 minute. Dilute the 100× concentrated HRP enzyme conjugate to a 1× working concentration with universal diluent (e.g., 10 μL of concentrate + 990 μL of universal diluent). Prepare immediately. 5. Prepare the 1× wash solution: Dispense 10 mL of 20× wash solution into 190 mL of distilled water (concentrated wash solution removed from the refrigerator may crystallize; this is normal. Allow to stand at room temperature until the crystals have completely dissolved before preparing). Procedure: 1. Remove the desired strips from the aluminum foil bag after equilibration at room temperature for 10 minutes. Seal the remaining strips in a ziplock bag and return to 4°C. 2. Sample addition: Add 100 μL of sample or standard of varying concentrations to the corresponding wells. Add 100 μL of universal diluent to the blank wells. Cover with a film and incubate at 37°C for 60 minutes. (Recommendation: Dilute the sample to be tested at least 1-fold with universal diluent before adding it to the ELISA plate. This will reduce the impact of matrix effects on the test results. The sample concentration should be multiplied by the corresponding dilution factor when calculating the final sample concentration. It is recommended to run replicates for all test samples and standards.) 3. Add Biotinylated Antibody: Remove the ELISA plate and discard the liquid without washing. Add 100 μL of Biotinylated Antibody Working Solution directly to each well. Cover with a film and incubate at 37°C for 60 minutes. 4. Wash: Discard the liquid and add 300 μL of 1x Wash Solution to each well. Let stand for 1 minute, shake off the wash solution, and pat dry on absorbent paper. Repeat this process three times (a plate washer can also be used). 5. Add Enzyme Conjugate Working Solution: Add 100 μL of Enzyme Conjugate Working Solution to each well. Cover with a film and incubate at 37°C for 30 minutes. 6. Washing: Discard the liquid and wash the plate five times as in step 4. 7. Adding substrate: Add 90 μL of substrate (TMB) to each well, cover with a sealing film, and incubate at 37°C in the dark for 15 minutes. 8. Adding stop solution: Remove the ELISA plate and add 50 μL of stop solution directly to each well. Immediately measure the OD value of each well at a wavelength of 450 nm. Calculating experimental results: 1. Calculate the average OD value of the standard and sample replicates and subtract the OD value of the blank well as a correction factor. Plot the standard curve of the four-parameter logistic function on double-logarithmic graph paper, with concentration as the horizontal axis and OD value as the vertical axis. 2. If the sample OD value is higher than the upper limit of the standard curve, dilute the sample appropriately and retest. Multiply the sample concentration by the corresponding dilution factor. |
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| Theory | This kit uses a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA). Sample, standard, biotin-labeled detection antibody, and HRP conjugate are sequentially added to microwells pre-coated with a Cytochrome P450 24A1 (CYP24A1) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by peroxidase (HRP) catalysis and to yellow by acid. The intensity of the color is positively correlated with the amount of Cytochrome P450 24A1 (CYP24A1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Cytochrome P450 24A1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Cytochrome P450 family 24, subfamily A, member 1 (abbreviated as CYP24A1), also known as vitamin D₃ 24-hydroxylase, is a member of the cytochrome P450 superfamily and is encoded by the CYP24A1 gene. It is a mitochondrial monooxygenase that catalyzes the 24-hydroxylation of calcitriol (1,25-dihydroxyvitamin D₃). Conditions associated with CYP24A1 include hypercalcemia, infantile dysregulation, and Brody disease. Pathways associated with this gene include metabolism and metabolism of fat-soluble vitamins. Gene ontology annotations associated with this gene include oxidoreductase activity and heme binding. An important homologue of this gene is CYP27A1. | |||||||||||||||||||||||||||||||||
| General Notes | 1. Strictly adhere to the specified incubation time and temperature to ensure accurate results. All reagents must be at room temperature (20-25°C) before use. Refrigerate reagents immediately after use. 2. Improper plate washing may result in inaccurate results. Ensure that all liquid in the wells is aspirated thoroughly before adding substrate. Do not allow the wells to dry out during incubation. 3. Remove any residual liquid and fingerprints from the bottom of the plate, as this will affect the OD value. 4. The substrate developer solution should be colorless or very light in color. Do not use substrate solution that has turned blue. 5. Avoid cross-contamination of reagents and specimens to prevent erroneous results. 6. Avoid direct exposure to strong light during storage and incubation. 7. Do not expose any reagents to bleaching solvents or the strong fumes emitted by bleaching solvents. Any bleaching agent will destroy the biological activity of the reagents in the kit. 8. Do not use expired products, and do not mix components with different product numbers and batches. 9. Recombinant proteins from sources other than the kit may not be compatible with the antibodies in this kit and will not be recognized. 10. If there is a possibility of disease transmission, all samples should be managed properly and samples and testing devices should be handled according to prescribed procedures. |
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| Storage Temp. | If the unopened kit is stored at 4°C, the shelf life is 6 months. | |||||||||||||||||||||||||||||||||
| Test Range | 0.156-10 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissuehomogenates, celllysates and otherbiologicalfluids |
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4.4 ★★★★★
Based on 30 reviews
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Product Reviews
★★★★★ 5
Great value
Size: Eufy B-18Packs
This 18-pack of replacement parts for the eufy L50 and L60 robot vacuums is very handy. It includes everything needed to keep the vacuum running smoothly, making maintenance easy and convenient. Great value for anyone who owns these models! better price than the OEM.
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Reviewed in the United States on March 30, 2026
★★★★★ 5
GOOD
Style: 28Pack Eufy Omni C20 Accessories
The kit includes several useful pieces and helped me replace accessories I already had worn out. I liked that everything came organized and it was easy to find each part. Some pieces feel sturdier than others, but overall it does the job well and it is convenient having everything together in one package.
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Reviewed in the United States on May 28, 2026
★★★★★ 5
Buena calidad precio
Style: 28Pack Eufy Omni C20 Accessories
Precio beneficio me parece un paquete bastante completo para hacer los mantenimientos. En cuanto a tamaño calidad y funcionamiento no hay queja (salvo por del atornillador que es como de mentira y se desarmo al tratar de aflojar el tornillo y no es de la medida correcta).
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Reviewed in the United States on November 8, 2025
★★★★★ 5
Excellent Long Term Value & Perfect Compatibility with Eufy Omni C20 Robot Vacuum
Style: 28Pack Eufy Omni C20 Accessories
This set is of great quality and value for the money. It fits perfectly with the vacuum and base station and includes extra tools like a small screwdriver if you need to make any repairs. I love how it functions and provides the user with an easy experience with all the maintenance tools in one kit, lasting for many months or even a year or two with the primary vacuum. I absolutely recommend this product. Everything you need!
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Reviewed in the United States on November 29, 2025
★★★★★ 4
Great value pack
Style: 28Pack Eufy Omni C20 Accessories
I've used each type of item in my vacuum and they all work just as well as the originals. The fit is precise and they last just as long as the original items. It's a great value pack and I love that I have so many extras in the house. I did ding one star because the included screwdriver is the wrong size to install the side brush. If you aren't paying attention during installation you could easily strip the screw. The other items are very easy to install.
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Reviewed in the United States on February 1, 2026
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