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Description
Mouse YAP1 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: Adherent cells should be gently washed with pre-chilled PBS, then trypsinized and harvested by centrifugation at 1000×g for 5 minutes. Suspension cells can be harvested directly by centrifugation. Collected cells should be washed three times with pre-chilled PBS and resuspended 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. 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 Yes Associated Protein 1 (YAP1) 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 Yes Associated Protein 1 (YAP1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Yes Associated Protein 1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Yes-associated protein 1 (YAP1), also known as YAP or YAP65, is a protein encoded by the YAP1 gene. It functions as a transcriptional regulator by activating the transcription of genes involved in cell proliferation and inhibiting apoptosis genes. YAP1 is inhibited in the Hippo signaling pathway, thereby enabling cellular control of organ size and tumor suppression. YAP1 was first identified through its ability to bind to the SH3 domain of the Yes and Src protein tyrosine kinases. It is a potent oncogene that is amplified in various cancers. It is a transcriptional coactivator whose proliferative and oncogenic activities are driven by its binding to the TEAD family of transcription factors, which upregulate genes that promote cell growth and inhibit apoptosis. | |||||||||||||||||||||||||||||||||
| 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, and other biological fluids |
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4.2 ★★★★★
Based on 25 reviews
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Product Reviews
★★★★★ 5
Cute, soft, comfy
Size: 6-12 Months Infant, Color: Navy
I wanted a leather sole shoe for my ten month old daughter for playing outside this summer so she could have protection for her feet without a hard “supportive” shoe inhibiting her ability to learn to balance or affecting her feet as the bones continue to develop.
These are perfect, they are soft and easy to put on. The elastic in the ankle holds it on without being too tight and it doesn’t dig into her skin at all. Provide protection and grip as she learns to walk and are not too bulky at all. I got the 9-12 month size and it has enough room to accommodate a chubbier foot than my daughter has, but still fits well and looks adorable.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on June 25, 2024
★★★★★ 5
Love these
Size: 18-24 Months Infant, Color: Grey
These are awesome! My 15-month old wore these everywhere on our trip to Mexico. He scuffed them up quite a bit, but other than cosmetics, they're holding strong through sand, sprinklers, and general gallivanting around. They are super easy to get on, which also makes them easy for him to pull off. But as long as he's busy running a muck, he forgets they're even on his feet.
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Reviewed in the United States on January 21, 2026
★★★★★ 4
Sizing is off, send me free pair!
Size: 3-4 Years Toddler, Color: Navy, Size: 3-4 Years Toddler, Color: Navy
Sizing is off. I’m very frustrated. I ordered it to Costa Rica thru shipping company in Miami, which takes about 3 weeks to deliver and + shipping fee that they apply (20$)
I spent time and checked the size guide. According to the size guide, I ordered “3-4 years old” which supposed to be 16cm from heel to toe) the pair I ordered ended up being 17cm!
My baby’s feet are 15cm, I was planning to get 16cm for him to grow into it, but 17cm is too big. The shoes it self looks good, the leather is very soft and nice. I’m not sure how durable they are. Ordering first time from this brand. I ordered 2 pairs of shoes from a similar brand and liked it. They just didn’t have the size I needed. Although, the leather the previous brand uses for shoes seems much thicker. Again, I don’t know how durable “Dooty Fish” but I like how soft on skin they are.
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Reviewed in the United States on August 21, 2024
★★★★★ 5
Wide and flexible
Size: 18-24 Months Toddler, Color: White
Very cute I love the width and flexibility of the shoe 18 to 24 month is a bit too big for my 18 month old and she generally wears a size 6 1/2 shoe. I’m going to try the smaller size.
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Reviewed in the United States on July 15, 2025
★★★★★ 1
Don’t buy
Size: 12-18 Months Infant, Color: Navy
Cute little shoes but I would not recommend people buy these. The elastic broke on both shoes within a month. And they cannot get even a tiny bit of water on them. (Which is impossible with a toddler) If they do get wet the bottoms become dangerously slick. I wanted to get nice little barefoot shoes that I could use as hand-me-downs for future kiddos but these were not it. Horrible quality and not worth the money.
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Reviewed in the United States on August 23, 2025