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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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4.2 ★★★★★
Based on 5 reviews
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Product Reviews
★★★★★ 4
Great for the entire family but expensive
So far so good. The brand of boxers we had been buying changed their style and construction so we needed to find an alternative. These have held up well so far to multiple weeks of frequent washing and wearing from the entire family. We did size up from the usual 6/M we normally buy and ordered large to prevent the legs from rolling up which worked out great. Our daughter likes the length and how soft the fabric is for wearing under skirts and gym shorts and liners stay put without moving or leaking as another reviewer mentioned. Our son says they are comfortable and fit well being just supportive enough without being constrictive. We all agree that there seems to be not enough fabric in the back as they tend to be much tighter across the backside than the front and that also causes the waist band to ride lower in the back than the front. These have better seam placement than some brands we tried with no seam across the lady bits and no seam down the middle of the back side so they are more much comfortable but the cost is also much higher and that is why we're only giving it 4 stars. These will stay in the rotation until we find something cheaper that both us and the kids like.
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Reviewed in the United States on January 24, 2026
★★★★★ 5
Pregnant lady review! Soft, comfortable, cool.
Color: 03-multicolour (4-pack), Size: Large
So soft, comfortable and cool. I'm pregnant and I've gained weight in my legs. I love wearing dresses but my thighs were rubbing together too much. These are perfect to wear under dresses. I also wear them under my bibs at work and it keeps me from getting overheated but I don't have to worry about whether or not my underwear show since they just look like shorts. I wish these came in maternity. I just keep them below my belly. I'll be buying more for sure.
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Reviewed in the United States on April 8, 2026
★★★★★ 5
I want to live in these
Color: 01-multicolour (4-pack), Size: Medium
I bought 4 different sets of new underwear. Unfortunately these came last. It’s unfortunate, because if they had come first I would have cancelled all my other orders or returned them and ordered more of these. I never want to wear anything else ever again; these are the comfiest undies I’ve ever owned, I want to live in them. I can wear them under pretty much anything except leggings or very tight pants (but let’s be honest, I don’t wear those anyway.)
They are incredibly soft and breathable, they wash well, don’t roll, keep my thighs from rubbing, are comfy to sleep in, and the colors were true to the photo.
Just a heads up, they are on the thin side and just a bit see through in the rear.
I’m 5’5” 140lbs and the mediums fit perfectly, they go about mid-thigh.
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Reviewed in the United States on July 30, 2025
★★★★★ 5
Extremely comfortable
Color: 02-multicolour (4-pack), Size: X-Large
These are by far the softest and most comfortable underwear I have ever owned. The leg portion absolutely does roll up as so many others have mentioned, however, I did not find that to be uncomfortable at all just annoying that they don’t stay in place.
I bought these specific underwear to see if it would help alleviate HS symptoms at all. They have definitely helped with wicking moisture away from the skin. More time will tell if it’s truly helpful for HS. For now, I will recommend these. They are thin, breathable, butter soft, wick moisture comfortable and I would buy again…maybe in a different style next time. I have washed them several times and they seem to be holding up rather well so that is also of supreme importance given the cost of the item.
Again, I would recommend this item and plan on purchasing more in a different style very soon.
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Reviewed in the United States on September 13, 2025
★★★★★ 3
Eaten by my Thighs
Color: 04-multicolour (4-pack), Size: X-Large
I wanted to love them. Sadly they have let me down.
Pros: natural breathable fabric , super soft and comfortable, cotton panty gussets for feminine health and panty liners will stick!! Yay:). And the colors! They are great!
But here’s the Con. And it’s the con of all cons.
THEY RIDE UP!! Eaten by my thighs. I’m 5’6” and in the general area of 175. I fluctuate depending on how many times I visit the bakery! :) I don’t have a thigh gap but it is not a super tight thigh squeeze. I seldom have thigh climb but these do. I tried two different sizes to see if that made a difference. Nope.
I was miserable at a recent theme park in these. I was wearing long heavy dresses and it was quite awkward to yank these legs down non stop.
They feel amazing and they have all the requirements
1. Soft
2. Comfortable and stretches with you
3. Breathable with nice thin fabric. Not bulky
4. Natural fibers
5. Great colors
If you are a person that is not blessed with Thunder Thighs, please order these. They are great! I wear them at home and to locations that I will not be doing a lot of walking at. I have washed them so many times and they look like new still.
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Reviewed in the United States on October 4, 2025