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Description
Mouse DKK3 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: 1. 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: 1. 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. 2. Plasma: Collect the specimen using EDTA or heparin as an anticoagulant. Centrifuge the specimen at 1000 × g for 15 minutes at 2-8°C within 30 minutes of collection. The supernatant can be assayed or stored at -20°C or -80°C, but avoid repeated freezing and thawing. 3. Tissue homogenization: Rinse the tissue with pre-chilled PBS (0.01M, pH 7.4) to remove residual blood (lysed red blood cells in the homogenate will affect the measurement results). Weigh the tissue and mince it. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1 g of tissue sample to 9 mL of PBS. The specific volume can be adjusted according to experimental needs and recorded. It is recommended to add protease inhibitors to the PBS) in a glass homogenizer and grind thoroughly on ice. To further lyse tissue cells, the homogenate can be sonicated or repeatedly frozen and thawed. Finally, centrifuge the homogenate at 5000 × g for 5-10 minutes, and the supernatant can be assayed. 4. 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 utilizes 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 Dickkopf Related Protein 3 (DKK3) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by HRP peroxidase and to yellow by acid. The intensity of the color is positively correlated with the amount of Dickkopf Related Protein 3 (DKK3) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Dickkopf Related Protein 3 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Dickkopf-related protein 3, also known as DKK3, is encoded by the DKK3 gene. This gene encodes a protein that belongs to the Dickkopf family. The secreted protein contains two cysteine-rich domains and is involved in embryonic development through interaction with the Wnt signaling pathway. Dkk3 is a negative regulator of β-catenin, and its downregulation may contribute to the activation of β-catenin signaling in cervical cancer. Therefore, Dkk3 could be used as a therapeutic gene targeting β-catenin. | |||||||||||||||||||||||||||||||||
| 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, tissue homogenates and other biological fluids |
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4.6 ★★★★★
Based on 14 reviews
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Product Reviews
★★★★★ 5
They are her favorites
Size: Medium (VALUE 2-Pack), Style: Beef, Size: Medium (VALUE 2-Pack), Style: Beef
We’ve had many dogs, many very destructive ones and wish I had these years ago. The hard toys get sharp edges, this toy is the best and yes is far better than the other red ones. My bulldog wants tug of war for 30 min to an hour every night. We’ve gotten tennis elbow, tendinitis, and pulled muscles holding onto this thing. She has tons of toys but 9pm and this is only option for her. We don’t get a choice. Easy to hold, well for a few min until your hand gets tired. First you one was about 3 years ago and she’s only on her third one. They last forever, easy to clean, and aren’t sharp edges. Seriously, better than any toy sold anywhere. My phone has hundred of pictures of Pickle holding this. She’s doing it right now. If you have a strong destructive dog that takes pride in destroying toys, ruin their confidence and get them this one.
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Reviewed in the United States on April 5, 2026
★★★★★ 5
Perfect Toy for Large Dogs
Size: Medium, Style: Cheddar Cheese
I don’t normally leave reviews, but this toy is stellar and is a new favorite for my dog! She is a large dog (70lbs cattle dog lab mix) and I accidentally bought a medium, but it actually is a great size for her! So far this stands up to her chewing, but I may need to update in a few weeks to speak for durability but it really does feel solid. Most toys don’t last an afternoon once she starts chewing, so this has already won in that regard. The shape is perfect as well. She can gnaw at it, squeak, play fetch, play tug. It bounces when she throws it around herself. The fact it smells like stinky cheese is just an added bonus for her. All around, a very well designed dog toy!
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Reviewed in the United States on December 27, 2023
★★★★★ 4
Boxer Baby’s Favorite
Size: Jumbo, Style: Beef
This toy is my sweet girl’s absolute favorite. It was the first thing we gave her when we brought her home, and she has been obsessed with it since. She brings it from room to room, inside and outside. I would highly recommend this toy, however, we have gone through six in one year. They’re not very durable, especially with our boxer mix, who is a very heavy chewer. I’ve gotten the red and the blue, and she will only take red, which is fine with me because the blue STINKS. The red stinks too, but less so. The squeak is very loud, but she genuinely loves it so much that it’s hard to be annoyed by it. I’d say give the toy a try, you may be unintentionally giving them their favorite thing ever lol.
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Reviewed in the United States on October 2, 2025
★★★★★ 1
Design Hazard
Size: Medium, Style: Bacon, Size: Medium, Style: Bacon
I have one of these in a larger version I picked up at the petstore. This one I do not recommend. There is a whole in the center where the pet bites down and my doggies' teeth kept getting stuck in it. If you've ever had a toy get stuck in your pups mouth, you've seen how thet go nuts trying to get it out.
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Reviewed in the United States on April 18, 2026
★★★★★ 5
Cane Corso approved
Size: Jumbo, Style: Beef
I have two Cane Corsos, One is 80 pounds the other is 120 pounds. They can seriously destroy some toys very easily. These chewies have actually lasted for over six months at this point, which is pretty much miraculous. I bought them some more just so I always had plenty of them laying around as this discourages them from getting into trouble, chewing things they shouldn’t be chewing. They walk around with these and their mouths all the time and even after all these months, the squeakers still work. These are really high-quality and if you have a dog that just destroys toy toy, I highly recommend these. Even though they’ve had these for months, they go back to them to chew on and play with , it’s also really funny because they walk around with one and hanging out of their mouth and it looks like they’ve got a stogie hanging out of their mouth. If these toys can keep the interest and survive being mauled by these two giant Destructo mutts, then they must be really, really tough. I highly recommend these for super chewers. These even a hold up to having one dog, grab each end and then play tug-of-war with it. These guys can Easily go through A cow huff every day, but these toys are still going strong
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Reviewed in the United States on August 10, 2024