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Description
Human Fetuin B ELISA KitProduct Specification Usage Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High precision pipette and gun 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: Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4 overnight, then centrifuge at 1000g for 20
Product Specification
| Usage |
Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High-precision pipette and gun 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: Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4℃ overnight, then centrifuge at 1000×g for 20 minutes, and take the supernatant, or store the supernatant at -20℃ or -80℃, but avoid repeated freezing and thawing. Plasma: Collect specimens using EDTA or heparin as anticoagulants and centrifuge them at 1000×g for 15 minutes at 2-8℃ within 30 minutes of collection. The supernatant can be tested or stored at -20℃ or -80℃, but repeated freezing and thawing should be avoided. Other biological fluids: Centrifuge at 1000xg for 20 minutes, remove the supernatant, and test. Pre-test preparation: 1. Remove the test kit from the refrigerator 10 minutes in advance and equilibrate to room temperature. 2. Prepare the standard gradient working solution: Add 1 mL of universal diluent to the lyophilized standard, let it stand for 15 minutes to completely dissolve, then gently mix (concentration is 20 ng/mL). Then dilute to the following concentrations: 20 ng/mL, 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 ng/mL, and 0 ng/mL. Serial dilution method: Take 7 EP tubes and add 500 μL of universal diluent to each tube. Pipette 500 μL of the 20 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 10 ng/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube serves directly as a blank well; there is no need to aspirate the liquid from the penultimate tube. See the figure below for details. 3. Preparation of Biotinylated Antibody Working Solution: 15 minutes before use, centrifuge the concentrated biotinylated antibody at 1000×g for 1 minute. Dilute the 100× concentrated biotinylated antibody to a 1× working concentration using universal diluent (e.g., 10µL concentrate + 990µL universal diluent). Prepare immediately before use. 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 enzyme conjugate are added sequentially to microwells pre-coated with fetuin B capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB converts to blue under the catalysis of HRP and to yellow under the action of acid. The intensity of the color is positively correlated with the fetuin B content in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Fetuin B ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Fetuin-B is a protein encoded by the FETUB gene in humans. The protein encoded by this gene is a member of the placenta family and belongs to the cysteine superfamily of cysteine protease inhibitors. Fetuins are involved in a variety of functions, including osteogenesis and bone resorption, regulation of insulin and hepatocyte growth factor receptors, and responses to systemic inflammation. This protein is likely secreted by cells. | |||||||||||||||||||||||||||||||||
| 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 | 0.31-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, and other biological fluids |
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4.9 ★★★★★
Based on 21 reviews
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Product Reviews
★★★★★ 5
Best Diaper Brand!
Size: Size 1, Unit Count: 96
Got these as a gift for my friend who just had a baby and she was thrilled! Never underestimate the simple gift of diapers to a momma with a newborn! Mother said she prefers these diapers because of their quality and softness. This is about the best price for high quality diapers you can find!
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Reviewed in the United States on May 12, 2026
★★★★★ 5
Great Product-Love These
Size: Newborn, Unit Count: 31
We have tried other name brand diapers over the years and people rave about other brands, but our experience with other brands is not good. Even the hospital doesn't use this brand and baby's onesie kept getting wet. They were folding their Pampers down at top for umbilical cord where Huggies have cut out for umbilical cord. Our personal experience is other name brand diapers seem to leak with our babies, and these stay snug and keep clothes and baby dry. When it says 100% leak proof and all-around blowout blocker it's factual for us and our babies. They are easy to use and durable. We have babies with sensitive skin and there is no irritation with these diapers.
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Reviewed in the United States on December 15, 2025
★★★★★ 5
Feat of diaper engineering
Size: Preemie, Unit Count: 30
I liked these well enough until my preemie had the biggest poo I’ve ever seen. That diaper probably weighed almost as much as he does and his white footie pajamas were untouched. Miracle. Worth every penny. Absolute legend.
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Reviewed in the United States on May 19, 2026
★★★★★ 5
Perfect thank you
Size: Newborn, Unit Count: 144
These baby diapers have been an excellent choice. They are very soft against the skin, which helps prevent irritation or chafing—even for babies with sensitive skin. Their absorbency is excellent, keeping the baby dry for several hours, both day and night. Furthermore, the fit is comfortable and secure, allowing the baby to move freely without any leaks. Without a doubt, they are practical, reliable, and provide peace of mind for parents. Highly recommended!
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Reviewed in the United States on May 3, 2026
★★★★★ 4
Outstanding, top of the line diaper!
Size: Preemie, Unit Count: 30
We normally use Huggies Little Movers with our 3.5 year old (nothing against them, they're also great diapers) but recently decided to give Swaddlers a try, and will not be going back. This is a top-of-the-line diaper and excels in all ways.
We use a size 7 though could *probably* get away with a size 6, and use about 4-5 diapers a day on average (not including his overnight diaper) depending on how much he's had to drink and how often he poops -usually once a day, but could be as many as 3 times, especially if he doesn't go one day.
The first thing you notice about these diapers is how soft they are. I mean, they just feel like they would be comfortable on a child, and since they're in them practically 24/7, that's important. The diaper is mostly white, and the overall designs are more muted than a lot of other diapers, with "Shilo and Friends" -an elephant and duck - on the front and back. I personally wouldn't mind something bolder, I don't think my child cares or even notices. There is a wetness indicator that goes down the length of the diaper, front and back. Nice to have I suppose for a caregiver who maybe doesn't change a lot of diapers, but at this age and stage, it's easy to know and tell. Like all Pampers diapers, they are scented. I happen to love the gentle powder-like scent, but it's a turn-off to some who describe it as too strong, and of course, if your child is sensitive to that, you may want to avoid Pampers diapers.
Inside, the top sheet liner keeps his skin dry, even when he wets heavily -which is usually - and does a good job of keeping poop from sticking to his bottom. The "blowout barrier" in the back also helps contain any messes. In terms of absorbency, I'm pretty amazed at how much they can hold. Being a little older, he tends to stay drier for longer periods and then pee a larger amount at one time, but the diaper almost always holds it in, and leaks are rare.
The one thing I wish that Pampers did do was have wider tapes, like Huggies, at least on the larger sizes, where kids are more active. It hasn't been a problem, so I don't really have a reason for saying that, other than that it seems like it would help with fit, but it certainly won't keep me from buying or using them in the future.
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Reviewed in the United States on May 18, 2026
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