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Description
Human WNT3A ELISA KitProduct Specification Usage Required experimental equipment: 1. Microplate reader (450nm) 2. High precision pipettes and pipette tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37C incubator 4. Distilled or deionized water Sample preparation and requirements: 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 and
Product Specification
| Usage | Required experimental equipment: 1. Microplate reader (450nm) 2. High-precision pipettes and pipette tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37°C incubator 4. Distilled or deionized water Sample preparation and requirements: 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 and mince the tissue. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1g of tissue sample to 9mL 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 collect the supernatant for analysis. Cell Lysis Buffer: Gently wash adherent cells with pre-chilled PBS, then trypsinize and collect the cells by centrifugation at 1000×g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash the collected cells three times with pre-chilled 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 freezing and thawing or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and collect the supernatant for analysis. Cell Culture Supernatant: Centrifuge at 1000×g for 20 minutes. Collect the supernatant for analysis, or store at -20°C or -80°C, but avoid repeated freezing and thawing. 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 enzyme conjugate are sequentially added to microwells pre-coated with a Wingless Type MMTV Integration Site Family, Member 3A (WNT3A) 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 amount of Wingless Type MMTV Integration Site Family, Member 3A (WNT3A) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Wingless Type MMTV Integration Site Family, Member 3A ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Wingless-type MMTV integration site family member 3A (WNT3A) is a protein encoded by the WNT3A gene. The WNT gene family consists of structurally related genes that encode secreted signaling proteins. These proteins are crucial in tissue homeostasis, embryonic development, and disease. Wnt3A sculpts a population of multipotent stem cells that give rise to neurons, muscle, bone, and cartilage in the trunk region. It instructs these multipotent stem cells to form muscle, bone, and cartilage progenitor cells rather than neurons. It also regulates the Notch pathway, controlling the segmentation clock required for normal trunk development. | |||||||||||||||||||||||||||||||||
| 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 | Tissue homogenates, cell lysates, cell culture supernatants, and other biological fluids |
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4.6 ★★★★★
Based on 13 reviews
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Product Reviews
★★★★★ 5
Dog toy
Terrific dog toy
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on October 23, 2025
★★★★★ 5
Durable tug toy that our pitbull absolutely loves
As soon as I pulled this toy out, our pitty saw it and immediately wanted to play. Tug is her absolute favorite thing to do, and she is crazy about this one. She constantly looks for anyone willing to play with her. That makes me really appreciate the grip loops because they keep our hands comfortable during the tug sessions making it easy to play with her.
Our house is also run by an 8 lb Chihuahua who is definitely the alpha. Believe it or not, our big pitty is scared of the little dog and just lets her steal the toy away. She just patiently waits and grabs it back the second the Chihuahua loses interest. The size is great as both dogs can get good bites on it for secure hold.
We have had it for a month now, and it is holding up perfectly without any real damage. Best of all, our she still enjoys it just as much as she did on day one. I really like that it is made in the USA from BPA and latex-free materials. It cleans up easily in the dishwasher and even floats when we take it down to the ponds.
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For anyone interested, the exact product I purchased was listed as:
Wagalicious, Tuggfinity, Lime
The ASIN for the version I received was:
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Reviewed in the United States on May 16, 2026
★★★★★ 4
Fun Toy for Large Dogs, but Hard to Grab from Flat Surfaces
I gave this to a large dog to see how it would hold up during play. He really enjoys it and stays engaged with it, especially during active play sessions. The material feels durable enough to handle a bigger dog’s strength. It’s bright and easy to spot, which helps when playing outdoors. It’s clearly designed for interactive play.
One thing I noticed is that it can be a little difficult for him to pick up off the ground. The shape doesn’t give much to grab onto when it’s lying flat. Once he gets it, he enjoys it, but getting started can take a second. It’s still a fun toy, just something to keep in mind. Works best when tossed or handed to them.
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Reviewed in the United States on May 5, 2026
★★★★★ 5
An absolute powerhouse for heavy chewers—survived the "same-day" test!
Finding a toy that can survive a large dog is usually an expensive game of trial and error. More often than not, new toys end up destroyed in the exact same day we buy them. However, the Wagalicious Tuggfinity Lime toy has completely defied expectations.
This large, lime green tug toy is built like a tank. It is the perfect size for larger breeds and has actually held up incredibly well to intense tugging and heavy chewing sessions. Instead of shredding into pieces within hours, it’s still going strong.
The best part is the price point. At around $15, the value is absolutely awesome. Usually, you have to pay a massive premium for "indestructible" style dog toys, but this one gives you top-tier durability without breaking the bank. If you have a big dog that demolishes everything, this is a must-have.
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Reviewed in the United States on June 4, 2026
★★★★★ 5
Dad approved
I bought this because my dogs do not want “gentle play.” They want something they can really grab, shake, and try to win. This has been a great tug toy for that. The material feels thick, the stitching has held up, and it has enough grip that I can keep control without it slipping. It is also easy to rinse off after backyard dirt and dog drool. This is the kind of toy that actually gets used instead of ending up ignored in the corner.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on June 5, 2026
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