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Description
Human XDH 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 the sample using EDTA or heparin as an anticoagulant and centrifuge it 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℃. However, 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 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 500uL of universal diluent to each. Pipette 500uL of the 10ng/mL standard working solution into the first EP tube and mix thoroughly to make a 5ng/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 xanthine dehydrogenase (XDH) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by HRP peroxidase catalysis and to yellow by acid. The intensity of the color is positively correlated with the amount of xanthine dehydrogenase (XDH) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Xanthine Dehydrogenase ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Xanthine dehydrogenase, also known as XDH, XO, or XAN1, is a protein encoded by the XDH gene. It belongs to the group of molybdenum-containing hydroxylases involved in purine oxidative metabolism. The enzyme is a homodimer. Xanthine dehydrogenase can be converted to xanthine oxidase by reversible sulfhydryl oxidation or irreversible proteolytic modification. Its deficiency can cause xanthinuria, potentially leading to respiratory stress syndrome, and may enhance influenza infection through an oxygen metabolite-dependent mechanism. Patients with lung adenocarcinoma tumors that express high levels of the XDH gene have poor survival. Exploiting the XDH protein has been exploited to target NSCLC tumors and cell lines with precision oncology. | |||||||||||||||||||||||||||||||||
| 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 | Serum, plasma, breast milk, and other biological fluids |
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4.5 ★★★★★
Based on 9 reviews
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Product Reviews
★★★★★ 4
Keep chewing boys!
Color: KING KONG
There is nothing that can't be destroyed. The nice thing about dog toys from this company, is that they last a long time. We have 3 dogs 🐕, 2 of them are aggressive cheers. Over time, these dog toys will develop some
sharp edges. When the dogs want to play, if they rub that toy on your ankle, it can remove skin. The toy is still intact, but becomes hazardous to the humans in the house.
That is when you go back to the Amazon site, and order new toys.
Our dogs love these, as they are a challenge to chew. And they do last a long time.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 13, 2026
★★★★★ 5
Our Pit loves them!!
Size: Medium, Style: Bone, Product Packaging: Standard Packaging
We are on our third one in 2 years for our Pitbull Terrier. She’s an extremely aggressive chewer and will shred even the most “durable” labeled toys. She gets bored with rope style toys and always alternates to this Kong rubber bone. She will gnaw on this thing for hours. After a few days of use, it does start to show wear and the rubber begins splitting, but we haven’t had to be concerned about chunks or anything like that coming off and choking her. Keeps her teeth nice and shiny while keeping her occupied for hours. When one wears out, we just order a new one. I hate spending big money on dog toys, but these are more affordable and worth the cost!!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on June 7, 2025
★★★★★ 5
Durable, no bounce or squeak
Size: Large, Style: Ball, Product Packaging: Standard Packaging
My Great Dane loves to carry his ball on walks, but with most balls, when dropped, they’d bounce and roll away. I found these balls that are very durable when he bites down on them, and when he accidentally drops them, they don’t bounce or roll far.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on June 5, 2026
★★★★★ 4
Super fun for the dog, but doesn’t last.
Size: Small, Style: Bone, Product Packaging: Standard Packaging, Size: Small, Style: Bone, Product Packaging: Standard Packaging
I just ordered the 3 rd one. The bad thing is that my dog destroyed it in 3 hours. Luckily she spits out all the small pieces of plastic. The good thing is, that she gets so much joy out of it, it makes her so happy and tired that I am a fan! Well worth 9$!
She is 25lb and I didn’t think she was an aggressive chewer before this toy. If your dog tends to eat all the plastic pieces , don’t get it.
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Reviewed in the United States on April 4, 2026
★★★★★ 3
See updated review; destroyed within 15 minutes
Size: Medium, Style: Bow Tie, Product Packaging: Standard Packaging
I'm bummed. My pup destroyed this within fifteen minutes. I thought because it was made by Kong, it would hold up a little better. I also have their black rubber toys and those are great!
Update: I didn't expect to be contacted by the seller but they reached out after seeing my review and offered a full refund which I received quickly. Truly outstanding customer service!!!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on December 2, 2025
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