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Description
DPD 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 200 ng/mL). Then dilute to the following concentrations: 200 ng/mL, 100 ng/mL, 50 ng/mL, 25 ng/mL, 12.5 ng/mL, 6.25 ng/mL, 3.125 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 200ng/mL standard working solution into the first EP tube and mix thoroughly to make a 100ng/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 Biotin-Antibody Working Solution: Centrifuge the concentrated Biotin-Antibody at 1000×g for 1 minute 15 minutes before use. Dilute the 100× concentrated Biotin-Antibody to a 1× working concentration with universal diluent (e.g., 10uL concentrate + 990uL universal diluent) and use on the same day. 4. Preparation of Enzyme Conjugate Working Solution: 15 minutes before use, centrifuge the 100x concentrated enzyme conjugate at 1000×g for 1 minute. Dilute the 100x concentrated HRP enzyme conjugate to a 1x working concentration with universal diluent (e.g., 10µL concentrate + 990µL universal diluent). Use the same day. 5. Preparation of 1x Wash Solution: Dispense 10mL of 20x Wash Solution into 190mL 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 them to 4°C. 2. Sample Addition: Add 50 μL of sample or standard of varying concentrations to the corresponding wells. Add 50 μL of universal diluent to the blank wells, followed by 50 μL of Biotin-Antibody Working Solution to each well. Cover with film and incubate at 37°C for 1 hour. (Recommendation: Dilute the sample to be tested at least 1-fold with universal diluent before adding it to the ELISA plate to minimize matrix effects. The sample concentration should be multiplied by the dilution factor when calculating the final concentration. It is recommended to run replicates for all samples and standards.) 3. Plate Wash: Discard the liquid and add 300 μL of 1x Wash Solution to each well. Let stand for 1 minute, then shake off the wash solution and pat dry on absorbent paper. Repeat this process three times (a plate washer can also be used). 4. Enzyme Conjugate Working Solution: Add 100 μL of enzyme conjugate working solution to each well. Cover with film and incubate at 37°C for 30 minutes. 5. Wash: Discard the liquid and wash the plate five times as in step 3. 6. Add 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. 7. Add 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. Calculation of experimental results: Result evaluation: 1. Calculate the average OD value of the standard and sample replicates and subtract the OD value of the blank well as a correction value. Plot a standard curve for 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 competitive enzyme-linked immunosorbent assay (ELISA). Sample, standard, biotin-labeled antibody, and HRP conjugate are sequentially added to microwells pre-coated with a universal deoxypyridinol/deoxypyridinoline (DPD) antigen. After incubation and washing, the assay 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 universal deoxypyridinol/deoxypyridinoline (DPD) content in the sample. The absorbance (OD value) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | All | |||||||||||||||||||||||||||||||||
| Synonym | General Deoxypyridinol;DeoxypyridinolineELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Competition Law | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Deoxypyridinoline (DPD), also known as D-pyrilinks, Pyrilinks-D, or deoxyPYD, is one of two pyridine crosslinks that provide structural rigidity to type I collagen in bone. It is measured in urine tests and used along with other bone markers such as alkaline phosphatase, osteocalcin, and N-terminal telopeptide to diagnose bone diseases such as postmenopausal osteoporosis, bone metastases, and Paget's disease. It is also useful in monitoring treatment with bone-active medications such as estrogens and bisphosphonates. | |||||||||||||||||||||||||||||||||
| 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 | 3.12-200ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, and other biological fluids |
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★★★★★ 5
My dogs are obsessed with this ball
Size: 2-pack, Style: Duo
Great quality! I have 2 boxers that love it and the ball is still intact
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Reviewed in the United States on January 31, 2026
★★★★★ 5
Not Scented, but Dog Approved
Size: 2-pack, Style: Duo
They don't smell like peanut butter or bacon. But our dogs still love them.
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Reviewed in the United States on August 31, 2025
★★★★★ 4
If they squeaked, they would be the perfect ball!
Size: 2-pack, Style: Duo
I bought our 15 month old Coton several Chuck-It toys on an Amazon deal. He is incredibly playful, mischievous, and ball/toy obsessed, especially for anything that makes noise. He can also be very destructive so I love how durable most of their products are - they’re among our longest lasting toys and worth every penny. We have yet to take them to the dog park, but they’re a perfect size for sharing in plat time. And they are keeping him busy at home - that’s a good thing. The scent is subtle but strong enough for him I guess because he loves gnawing on them. If they had squeakers, they would be absolutely perfect. Hopefully Chuck-It will see an opportunity for that down the road!
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Reviewed in the United States on May 28, 2026
★★★★★ 5
Most durable fetch balls I know of.
Size: 2-pack, Style: Duo
These balls are the biggest balls of them all. Well, not the biggest, but the best. Every other ball she would chew up in short order and I'd be throwing her a floppy piece of rubber. She just wants to fetch, but if it's not bouncing, she can't catch it on the bounce. Got these balls in July and no damage to them. I just ordered another pair, but only because the other two somehow got stuck up in trees. We have some very old and very large oak trees so not so easy to retrieve. Hope they somehow drop someday. But in the meantime, Lassie will be happy with her new ones. Very highly recommended!
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Reviewed in the United States on October 28, 2025
★★★★★ 5
Works great for scent work! Bounce is great for air retrieve!
Size: 2-pack, Style: Duo, Size: 2-pack, Style: Duo
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Reviewed in the United States on August 6, 2025