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Description
Human VIL 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 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 villin 1 (VIL) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by peroxidase (HRP) catalysis and to yellow by acid. The intensity of the color is positively correlated with the amount of villin 1 (VIL) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Villin 1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Villin 1 (VIL), also known as VIL1, is a protein encoded by the VIL1 gene. This gene encodes a member of the calcium-regulated actin-binding protein family. This protein represents a major component of the brush-border cytoskeleton, playing a role in the capping, severing, and bundling of actin filaments. Two mRNAs, 2.7 kb and 3.5 kb, have been observed, each utilizing an alternate polyadenylation signal present in the terminal exons. It plays a role in actin nucleation, actin filament bundle assembly, and actin filament capping and severing. It regulates intestinal epithelial cell morphology, cell invasion, cell migration, and apoptosis. It protects against dextran sodium sulfate (DSS)-induced apoptosis in gastrointestinal epithelia. It appears to regulate cell death by maintaining mitochondrial integrity. It enhances hepatocyte growth factor (HGF)-induced epithelial cell motility, chemotaxis, and wound repair. | |||||||||||||||||||||||||||||||||
| 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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4.8 ★★★★★
Based on 21 reviews
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Product Reviews
★★★★★ 5
DERM APPROVED!
I am obsessed with this product!! This feels very luxurious on my skin and does not pill like other products I have had in the past. I have noticed a significant improvement in the background redness in my skin. As someone who works in derm and values good ingredients and products that actually work... I can confidently say that this is the best OTC azelaic acid product I have tried out. It sits very nicely under my moisturizer and does not dehydrate my skin like other azelaic acid products have done in the past. HIGHLY RECOMMEND
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Reviewed in the United States on May 10, 2026
★★★★★ 5
A Game Changer for Sensitive Skin
I use this serum every single day, and it has become one of the most important products in my skincare routine. I have very sensitive skin and cannot tolerate stronger active ingredients such as retinol or high-strength acids, so finding something effective but gentle has been incredibly important.
This serum has helped calm redness, reduce inflammation, and noticeably improve my skin’s overall appearance and comfort. After dealing with a prolonged inflammatory skin condition, this product made a significant difference and helped restore my skin barrier.
The formula feels soothing, hydrating, and non-irritating. If you have sensitive or reactive skin and are looking for a gentle product to help with redness and uneven texture, I highly recommend giving this serum a try.
Photos attached: The green bottle is the product and the second photo includes my favorite additions.
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Reviewed in the United States on May 13, 2026
★★★★★ 5
Perfect for Acne-Prone Skin!
This serum has honestly been a great addition to my routine for redness and post-acne marks. The texture is lightweight, hydrating, and doesn’t feel sticky or heavy on my oily/acne-prone skin.
I have sensitive skin with a focus on barrier protection, and this didn’t irritate my skin at all. It layers beautifully with the rest of my skincare and leaves my skin looking calmer and smoother over time. AND, it doesn’t have a smell!!
Definitely one of my favorite azelaic acid serums I’ve tried.
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Reviewed in the United States on May 18, 2026
★★★★★ 5
6-Month Update: Still a staple for sensitive, acne-prone skin
Size: 3.38 Fl Oz (Pack of 1), Size: 3.38 Fl Oz (Pack of 1)
I originally reviewed this ampoule when I first started using it, and now after about six months of consistent use (morning and night), I feel confident giving a more long-term update.
I have sensitive, acne-prone, combination skin, and this has remained one of the most reliable products in my routine. The texture is still one of my favorite things about it — very lightweight, absorbs quickly, and layers well under moisturizer and sunscreen without pilling.
Over time, I’ve noticed:
Less visible redness
More balanced oil production in my T-zone
Fewer irritation flare-ups when introducing new products
No clogged pores or breakouts from this formula
It doesn’t feel heavy or sticky, even in humid weather. I’ve continued repurchasing and even keep it on subscription because it consistently performs without causing sensitivity.
If you’re looking for a gentle, soothing centella product that works well for long-term barrier support, this has proven to be a dependable option in my experience.
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Reviewed in the United States on December 9, 2025
★★★★★ 5
Healed my barrier so fast
Size: 3.38 Fl Oz (Pack of 1)
I received this ampoule yesterday and I'm already leaving a 5 star review after 2 uses because it just worked so well. I bought it because I've recently incorporated the Timeless 20% Vitamin C into my routine and that + my tretinoin made my skin drier than I've ever felt it before, even gently swiping a cotton pad with micellar would sting on some parts of my face. As soon as this ampoule arrived I applied it to clean skin and layered squalane on top (my moisturizer was burning as well.) This product did not sting my face AT ALL. It instead felt immediately soothing and hydrating. When I woke up this morning almost all of my redness was gone, my skin didn't feel tight or irritated, and my skin tone looked so even and glowy. I never thought a product, especially such a simply formulated one, could heal my barrier so quickly but that's exactly what it did and I can already tell it's a repeat buy. If you have redness prone sensitive skin and/or you use several high strength actives, this ampoule is a must have and it might be the key for anyone struggling to tolerate high strength retinoids. I got the largest size bottle for $15 on sale and it's seriously one of the most affordable and effective skincare products I've ever had in my routine.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 30, 2026
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