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Description
Human 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. Cell Lysis Buffer: Gently wash adherent cells with ice-cold PBS, then trypsinize and collect cells by centrifugation at 1000×g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash collected cells three times with ice-cold PBS and resuspend in 150-200 μL of PBS per 1×10^6 cells (it is recommended to add protease inhibitors to the PBS; if the cell count is very low, reduce the PBS volume appropriately). Disrupt the cells by repeated freeze-thaw cycles or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and remove the supernatant for analysis. 5. Cell Culture Supernatant: Centrifuge at 1000×g for 20 minutes. Remove the supernatant for analysis or store at -20°C or -80°C, avoiding repeated freeze-thaw cycles. 6. 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 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 | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human 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 | 0.312-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.0 ★★★★★
Based on 16 reviews
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Product Reviews
★★★★★ 5
It's worth it.
Size: Size 7, Unit Count: 44, Size: Size 7, Unit Count: 44
I would like to provide feedback regarding a recent diaper experience. My toddler, who has a tendency to remove his diaper, has previously used Parent's Choice products since birth without issue. However, I have recently encountered a problem with both their standard diapers and Pull-Ups, specifically concerning the coverage in the gluteal region. This has led to the diaper riding up and causing a significant rash due to friction.
I am pleased to report that after two hours of use, the current diaper has exceeded my expectations. My son has not removed it, and the coverage in the gluteal area is excellent. I will provide an update should any leakage occur. I also appreciate the baby powder scent and the overall quality of the product. While the price point is slightly higher than I typically prefer for diapers, my son is approaching three years old and will soon be transitioning to potty training.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 3, 2026
★★★★★ 5
Movement freedom and quick changes: The ideal diaper for active babies
Size: Size 5, Unit Count: 56, Size: Size 5, Unit Count: 56
360° Flexible Fit: The standout feature is the 360° stretchy waistband that adapts perfectly to the baby's body, preventing gaps and allowing full, unrestricted movement.
• Easy On and Off: Being a "Pull-On" style, it slides up in a second. For removal, it features the practical "e-z off" tabs on the sides that tear easily for a hygienic and fast change.
• Designed for Active Babies: The packaging highlights an All-Around Fit specifically engineered for dynamic little ones who need protection that keeps up with them.
If your baby has started moving and diaper changes have become a 'wrestling match' with traditional tabs, Pampers Cruisers 360° are the ultimate solution. They offer the peace of mind of Pampers protection with the convenience of pull-up underwear. A great investment for your child's exploration stage.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 1, 2026
★★★★★ 4
Great swaddlers alternative when the Velcro became an issue
Size: Size 3, Unit Count: 136
I bought these because my daughter (12 months) started undoing the Velcro on her diapers, even through her clothes. Finding her soaking wet enough times led me to look for a solution. I really like these and will enjoy them until she finds her way out of them. They’re just as absorbent as Swaddlers and the overnights, and she never gets a rash in them. Only downside is the blowout barrier on the legs is much smaller, so she does have some blowouts out the side. The Swaddlers has more material there to prevent blowouts. These are also cheaper than Swaddlers and overall great quality. Soft and absorbent and I don’t have to worry about her even through a 13 hour sleep at night. That thing will weigh 5lbs when I pull it off her but it doesn’t leak! These are going to be our go to diapers as long as she doesn’t learn to pull them off.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 26, 2026
★★★★★ 5
Great fit and very convenient for active babies
Size: Size 4, Unit Count: 64, Size: Size 4, Unit Count: 64
These diapers are a great option, especially for active babies. The 360 design makes them easy to pull on and off, which is really convenient during quick changes.
They fit well without feeling too tight and provide good coverage. Absorbency is solid, keeping things dry for a good amount of time. The material feels soft and comfortable on the skin.
I also like how easy they are to dispose of after use. Overall, a reliable and convenient choice for everyday use.
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Reviewed in the United States on April 8, 2026
★★★★★ 5
What a wonderful treat, loved by our very fussy dog!
Our very fussy Havanese absolutely LOVES these sticks! I had to find a new product after the USA-made chicken wrapped rawhide sticks were no longer available. It is very difficult to find a quality product made in the USA & while these are made in Cambodia, they seem to be made under very strict standards. I was also impressed that this co. claims it has never had a recall of their products. I also relied on the many, many excellent reviews of past customers. Our Teddy gets a stick every morning & then will wait by the pantry to get an additional 1 or 2 or more throughout the day! He usually does not eat the rawhide, just chews it up a bit. He is in love with these!! He also has a sensitive stomach & these have never bothered him one bit. His teeth are also quite clean from using these!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 18, 2026
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