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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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4.4 ★★★★★
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Product Reviews
★★★★★ 5
Very solid!
Style: Corded
Really solid product for the price. Feels heavy and well made. Instructions are very clear and simple. This is why I buy Wen products!
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Reviewed in the United States on May 28, 2026
★★★★★ 4
Great budget multi tool
Style: Corded
Nice little rig that fits the bill for occasional hobby use. The motor is powerful enough to polish and cut without bogging down. Bits attach securely and easily. Light and easy to use. The price is great for this multi tool. Recommended
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Reviewed in the United States on December 22, 2025
★★★★★ 5
Favorite DIY Sidekick
Style: Corded
Bought this for one task: grind away an interfering piece of metal in my door’s strike plate. I was unhappy about spending $20 on another DIY tool for one tiny specific purpose but to my pleasant surprise, I haven’t stopped using this dremel since.
I’ve cut and sanded wood projects, trimmed custom cabinet panels, smoothed jagged hole edges, and more. I hardly ever turn the dial past 4 to find a speed I’m comfortable with. It’s very powerful and gives great results. I bought a cheaper one that busted on the first try and I’m so happy this was my next choice. It’s handled all the fixer-upper challenges I’ve asked of it.
The starter kit is a little sparse on the good stuff like saw blades and washers but very helpful. The case is convenient. My only gripe is I have to be intentional about placement to get it to close but the cord and extension tool is so long and useful that it makes up for it. It’s a little bulky but comfortable to hold and I’ve been able to reach all the narrow corners and tight spaces I’ve needed.
My next project is to cut away some protruding screw threads and my immediate thought was to reach for the WEN rotary. For the amount of value I’ve gotten out of it, this thing has paid for itself a few times already.
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Reviewed in the United States on December 8, 2025
★★★★★ 5
Perfect rotary tool for light or heavy project with power.
Style: Corded, Style: Corded
Polishing is my main purpose for using rotary tool. I've had hardell and dremmel before. Heavy use of these tool they get hot and wear out fast. Wen rotary tool is better. It's heavy duty, twice as big, 1.5 lbs, it has big motor and variable speed without the noise.
It's perfect for hobbies and for 3in polishing or sanding pad. It's polarized electric plug in. The attachments came with it is superior than any rotary tool I've used. Its a solid product. I can already tell undoubtedly this is a good product if not the best. The other rotary tool have the lock chuck button opposite of the power button which premarturely damages internal mechanism accidentally use the lock button while still spinning. Wen is power button is switch away from chuck lock button. It's not too big nor too small to use. The life expectancy for Wen rotary tool will likely be three times longer than dremmel or hardell in my guess because of its design.
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Reviewed in the United States on February 13, 2026
★★★★★ 5
Love this thing. But, love the company even more!
Style: Corded
This is my 1st rotary tool ever. So excited because the things you can do are limitless.
I received my tool and only used it 3 times. I was so happy I bought this but then the power switch broke, oddly.
I said well, you get what you pay for right? Well, I called the phone number listed here on Amazon, and they answered. I briefly explained what happened, and just asked if I could send them the main unit for replacement. The customer service person asked for my details, order number, etc.
Then within a few minutes, he said, Sir, we're just going to send you a new one, if that's okay. I was like huh? I said, I know it's been 8 months and that really would be awesome because I love it!
He said we stand behind all of our products and it's the least we can do, and we apologize for any inconvenience.
I was like huh? Lol.
It really is hard to find service like this these days. To be so excited about a 20 dollar tool, that you've always wanted but just never got around to getting, spending 20 bucks and not only getting a fantastic tool, but getting the authentic service we all deserve, priceless!
Thank you so much Wen and Amazon! Hope this helps others that are interested in this really useful and fun tool.
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Reviewed in the United States on November 12, 2023