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Description
Human eIF4E 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 20 ng/mL). Then dilute to the following concentrations: 20 ng/mL, 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 ng/mL, and 0 ng/mL. Serial dilution method: Take 7 EP tubes and add 500 μL of universal diluent to each tube. Pipette 500 μL of the 20 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 10 ng/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube serves directly as a blank well; there is no need to aspirate the liquid from the penultimate tube. See the figure below for details. 3. Preparation of Biotinylated Antibody Working Solution: 15 minutes before use, centrifuge the concentrated biotinylated antibody at 1000×g for 1 minute. Dilute the 100× concentrated biotinylated antibody to a 1× working concentration using universal diluent (e.g., 10µL concentrate + 990µL universal diluent). Prepare immediately before use. 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 capture antibody against eukaryotic initiation factor-4A (eIF4E). 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 eukaryotic initiation factor-4A (eIF4E) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human eukaryotic initiation factor-4A ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Eukaryotic initiation factor 4E (eIF4E) is a protein encoded by the eIF4E gene. It is involved in directing the ribosome to the cap structure of the mRNA. It is a 24 kDa polypeptide that exists both in its free form and as part of the eIF4F preinitiation complex. Almost all cellular mRNAs require eIF4E for protein translation. The eIF4E polypeptide is the rate-limiting component of the eukaryotic translation machinery and is involved in the mRNA-ribosome binding step of eukaryotic protein synthesis. | |||||||||||||||||||||||||||||||||
| 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 | 0.312-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates, cell lysates, and other biological fluids |
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4.4 ★★★★★
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Product Reviews
★★★★★ 5
Great shoes
Size: 13, Color: Black/White
Great shoes, bought for my grandson's birthday, he was very pleased with them. I'm very pleased that they came in many sizes.
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Reviewed in the United States on May 19, 2026
★★★★★ 5
Great Value for the Money!
Size: 10.5, Color: Black/White
Pleasantly surprised at this price point! Comfy, sturdy and a way better sole the Sketchers!
Amazon products in general have been really good quality at affordable prices!
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Reviewed in the United States on May 3, 2026
★★★★★ 1
Size not as expected
Size: 11.5, Color: Grey
Ordered same size as I always do. Way too large. Size listed was off by a long shot
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Reviewed in the United States on May 22, 2026
★★★★★ 4
Lightweight and True to Size — Best for Casual Use
Size: 9.5, Color: Navy, Size: 9.5, Color: Navy
It is nice to see the Amazon Essentials brand expand from household essentials to apparel and shoes.
I ordered the Amazon Essentials Men's Lightweight Knit Running Sneakers. The product is made in Vietnam and comes in a shoe box, which is nice. I've noticed that quite often, inexpensive shoes are delivered just in a plastic bag. The sneakers are ready to use and have no odors from glue, organic solvents, etc., which is common nowadays.
I wear a size 9W or 9.5. These sneakers are 9.5 and feel like a typical 9.5 size, without surprises. The product is advertised as lightweight, and it is true. Each shoe weighs below 0.6 lb (265 grams). Inside is soft memory foam; the shoes sit on the foot well, snug enough and comfortable, and are easy to put on and take off.
They seem like a good fit for spring and summer as casual footwear or for light gym use. The sole is simple and not designed for performance or high-impact activities. Overall, a solid, reasonably priced, comfortable everyday option, though not intended for serious running or training.
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Reviewed in the United States on April 15, 2026
★★★★★ 5
Budget buy while being as comfy and stylish as an expensive brand!
Size: 9.5, Color: Navy, Size: 9.5, Color: Navy
My 14 year old had been running his own business since he was 12. He does lawn care and snow removal. His go to shoes when it's too hot for his work boots has always been off brand hey d*de style ones which are terrible for that kind of work as they just don't have the ankle support.
He's also had some Orthopedic arch supports custom made for him through the DR in the last year that he needs to use in whatever shoes he's wearing at the time. So I've been on the search for a simple pair of slip on shoes that he could wear while working long hours around our community.
He's sent me this picture tonight before heading out. He said "I didn't feel like putting my arch supports in and these are so comfy that I don't even notice I don't have them!"
They are sharp looking shoes that really stand out as of they were A high end pair! It's great that they can be an every day style or could be more of a sport shoe as they have the support and padding for both options.
In our case they likely be green with in a day or two and it'll be interesting to see how well they wipe off between his work days. I can update this review once he's worn them a bit more.
For now they have been perfect for versatility, size was true to size (I did order 1/2 size up, but only did that because he's growing so fast!) and they are definitely a great fit to a budget!!!!
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Reviewed in the United States on May 8, 2026