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Description
Human WNT5A 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 conjugate are sequentially added to microwells pre-coated with a Wingless Type MMTV Integration Site Family, Member 5A (WNT5A) 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 Wingless Type MMTV Integration Site Family, Member 5A (WNT5A) 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 5A ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Wingless-type MMTV integration site family member 5A, also known as Wnt-5a protein, is encoded by the WNT5A gene. Disorders associated with WNT5A include Robinaud syndrome, autosomal dominant 1, and autosomal dominant Robinaud syndrome. Pathways associated with this gene include the canonical Wnt pathway and CDK-mediated phosphorylation and clearance of Cdc6. Gene ontology annotations associated with this gene include DNA-bound transcription factor activity and protein domain-specific binding. An important homolog of this gene is WNT5B. | |||||||||||||||||||||||||||||||||
| 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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4.9 ★★★★★
Based on 12 reviews
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Product Reviews
★★★★★ 5
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After the second day I bought another bottle. Just checked out as a matter of fact. Usually, even with my 3 separate moisturizing steps at night, my skin will have lost hydration the next morning. Even with a final layer of byoma oil it would feel like it pretty much evaporated out of my skin overnight. Idk what magic is in this stuff but I woke up today and it felt like I had JUST finished my nighttime skin care. Excellent under makeup..hell makes makeup look better. Worth every single penny.
Note: It is my experience that this tingles my skin a little bit. It's mild but it's there. The ingredients don't seem to contain anything that my skin would be sensitive to so it may just be in conjunction with another product in my routine. It's not unpleasant, just unexpected.
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Reviewed in the United States on February 27, 2026
★★★★★ 5
Amazing!
This sponge is amazing for applying the JI Pressed powder! I had been using a cheap sponge before but then decided to try this and it applies soooo much better. I think it’s because this sponge doesn’t absorb the makeup into it but instead releases it onto your skin for better coverage. It’s also incredibly silky smooth and allows you to smooth out and blend the makeup well. I don’t like using brushes so this is the perfect alternative!
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Reviewed in the United States on September 2, 2024
★★★★★ 5
Best makeup sponge
I use compact powder. Makeup goes on smooth and the sponge picks up enough product to give great coverage without multiple applications.
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Reviewed in the United States on May 18, 2025
★★★★★ 5
Absolutely amazing! Must try.
I absolutely love this sponge!, it is so dense;
it uses such a small amount of makeup but applies like a full coverage. I am using much less of my mineral makeup powder thanks to this sponge; plus it is easy to wash and dries quickly. Totally worth a try!
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Reviewed in the United States on March 11, 2024
★★★★★ 4
*update* Maybe(?) Perfect for those who hate eggs
Edit: Update
It is with a heavy heart Im updating this review. When I said this sponge stains, I mean it really stains. Now to be fair it is marketed as a powder sponge and I use it with liquid so this my be entirely on the user (aka me) but it's like I can never get it clean. Between my repeated washing and the overall product build up the performance of this sponge went downhill quickly. The finish of my makeup suffered and the flocking took a beating. For what it's worth the sponge has not fallen apart and is still in one piece. But I get more of a 4 star finish than a 5 star finish.
I've ended up using a large fluffy brush to buff out my foundation to get the desired result. I also ended up repurchasing the flocked flat ended egg from RT to replace this although at the moment I am still stubbornly using this sponge (and also avoiding doing my makeup)
I do believe a lot of my issues are do to using this sponge with liquid foundation instead of power and that is on me. I'm finding it so very difficult to clean and the RT flocked egg cleans up so much easier. I don't hate this sponge but my lovefest has definitely taken a turn to more of a likefest. I am going to keep using (because I am stubborn like that) and see how long it takes to turn this relationship into a hatefest or fall apart completely, whichever happens first. I will most likely update again at that time. At this moment I am changing my review from 5 star to 4 star. I will be keeping my original review below.
*original review*
Yo I LOVE this thing. Ok here's the deal, I don't like the beauty blender, in fact I kinda hate it. Yea, I said it, I'm not on board with bouncing an egg all over my face, I think it's tedious and inefficient.
I had at one time a flat teardrop shaped sponge that was my hands down favorite application tool for foundation. I used that thing until it literally fell apart. I tried to find a replacement but the store I had originally gotten it from no longer carried it so I kept using that ratty old sponge until it was missing chunks.
I tried some other flat sponges but none worked as I wanted so I went back to brushes but I really hate cleaning foundation from brushes so I was pretty much hating my life.
Then I found a flocked egg. Now I'm not a fan of the eggs but I was intrigued by the flocked texture and it had a nice flat edge so I tried it and it was almost as good as my old dearly missed flat teardrop sponge. I still was not a fan of bouncing an egg around on my face but I used this until the flocking wore off (about 6 months)
Well, I saw this listing for a flat flocked sponge and I pounced on it. I am so incredibly deliriously happy with this little sponge. I love that it's flat and not an egg. I love the flocking. I love how it fits in my hand. I love love the finish I get with my foundation. I love it. Like I need to order a dozen more so I never run out and I'm trying to break my hoarding tendencies but for this tool I want to hoard.
So the bad... It does stain easily. I wet my sponge before use and give it a quick clean after use. My old flocked egg would clean right up but this has stained on the first use. I also don't know how long this will hold up so I hope to update at a later time about that.
Overall I couldn't be happier with this sponge.
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Reviewed in the United States on February 10, 2021