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Description
Mouse YWHAe 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, biotinylated detection antibody, and HRP conjugate are sequentially added to microwells pre-coated with a capture antibody against Tyrosine 3/Tryptophan 5 Monooxygenase Activation Protein Epsilon (YWHAe). 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 Tyrosine 3/Tryptophan 5 Monooxygenase Activation Protein Epsilon (YWHAe) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Tyrosine 3/Tryptophan 5 Monooxygenase Activation Protein Epsilon ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | 14-3-3 protein epsilon (YWHAE) is a protein encoded by the YWHAE gene. This gene product belongs to the 14-3-3 protein family, which mediates signal transduction by binding to phospholipid-containing proteins. This highly conserved protein family is found in both plants and mammals, and this protein is 100% identical to its mouse homolog. It interacts with CDC25 phosphatase, RAF1, and IRS1 proteins, suggesting a role in various biochemical activities related to signal transduction, such as cell division and regulation of insulin sensitivity. It is also thought to be involved in the pathogenesis of small cell lung cancer. | |||||||||||||||||||||||||||||||||
| 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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4.8 ★★★★★
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Product Reviews
★★★★★ 5
Trusted reliable brand
Size: 5 Liter, Style: 1 Pack, Configuration: 5W-30
I recently used LIQUI MOLY Molygen New Generation 5W-30 for an oil change and overall I’m very impressed with the quality. The oil feels like a premium product and the difference in engine smoothness was noticeable almost immediately after the change. The engine runs quieter and smoother, especially during cold starts. This oil uses advanced additive technology designed to reduce friction and protect internal engine components, which helps improve efficiency and extend engine life. It also helps prevent deposits and keeps the engine cleaner over time. ()
Another thing I like is that it maintains stable viscosity even under high temperatures and heavy driving conditions. It provides excellent lubrication and wear protection while also helping improve fuel efficiency. The oil is also compatible with modern engines, including turbocharged applications, this is a high-quality synthetic motor oil that provides excellent protection and performance. If you’re looking for a reliable oil that keeps the engine running smoothly and helps reduce wear, this is definitely a great option and worth the price.
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Reviewed in the United States on March 7, 2026
★★★★★ 5
The best thing you can use in a Tahoe! Period!
Size: 5 Liter, Style: 1 Pack, Configuration: 5W-30
This is like liquid gold for your vehicle,it's molecular make up,is far superior to any oil out there,as it gets hot,it's molecules bind to every component inside your engine and attaches itself to the metal and fills in the stress points,all points really,it basically soaks into the metals,and is by far the most long lasting and does not break down like other oils,I get 10k off one oil change and I have 273,000 miles on my 03 Tahoe and this stuff is incredible I don't have any ticks,clicks,knocks, Nothing,not on fire up,cold start,my truck runs about five degrees cooler,I get a couple more miles to the gallon and my motor purrs like new,I could probably go 12-15k between oil changes,but I change it with the filter at 10K a new AC Delco pf46e and a 5-1/4 qt jug of liquid moly 5-30 full synthetic a new crush washer and I'm good for 10,000 miles Easily.
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Reviewed in the United States on May 17, 2026
★★★★★ 5
Smooth, Quiet, and Reliable Protection
Size: 5 Liter, Style: 1 Pack, Configuration: 0W-20
I recently switched over to this oil and it’s honestly one of the best upgrades I’ve made for my 2018 Mazda 3 Hatchback 2.5L. The 5 liter jug is the perfect amount for a full change, and I really like knowing it’s a high-quality fully synthetic formula that gives my engine the protection it deserves.
The first thing I noticed was how much smoother cold starts became—no more of that rough, clunky sound in the mornings. The engine feels quieter overall and runs more efficiently, like it’s not straining as much. I’ve also seen fewer deposits and buildup, which gives me peace of mind about long-term wear.
Another thing I appreciate is that it’s designed with modern engines in mind—safe for turbocharged setups and catalytic converters. It also blends easily with other oils, which makes topping off simple if I ever need to.
Highlights for me:
Excellent cold-start protection, even on chilly mornings.
Noticeable improvement in smoothness and reduced engine noise.
Works well with today’s engines, including turbos.
Easy to mix when topping off.
Overall, this oil gives me confidence that my engine is well-protected and performing at its best. Definitely feels like a premium choice that’s worth it.
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Reviewed in the United States on August 17, 2025
★★★★★ 5
Great Stuff!
Size: 5 Liter, Style: 1 Pack, Configuration: 5W-30
Purchased for my husbands F-150 Truck , his engine was making a weird noise and he thought he was going to have to get a new engine but a friend recommended to change his oil and use this one. Boom, it worked! Couldn't believe it cleaned the engine out and the noise went away. Engine seems to be working fine now. Will definitely use again!
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Reviewed in the United States on May 13, 2026
★★★★★ 5
Super nice and smooth, so much power! Car feels nimbler. Worth the price
Size: 5 Liter, Style: 1 Pack, Configuration: 5W-50
I normally don't buy such expensive engine oils (normally buy cheap diesel truck oils add in MoS2). However, this one was well worth the purchase. Engine feels substantially smoother and quieter. After running for a while, the car feels a lot nimbler. More power, and as if it simply wants to keep moving rather than whining. It glides like a magic carpet
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Reviewed in the United States on January 17, 2026