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Description
Human MIEN1 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 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 Migration and Invasion Enhancer 1 (MIEN1) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by HRP peroxidase and then to yellow by acid. The intensity of the color is positively correlated with the amount of Migration and Invasion Enhancer 1 (MIEN1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Migration And Invasion Enhancer 1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Migration-invasion enhancer 1, also known as MIEN1, is an enzyme encoded by the MIEN1 gene. It is a novel gene located next to HER2/neu in the chromosome 17q12 amplicon and has been shown to enhance the migration and invasion of prostate cancer cells. It is found to be abundantly expressed in breast tumor tissue and functions as a key regulator of tumor cell migration and invasion, promoting systemic metastasis. It increases cell migration by inducing filament formation at the leading edge of migrating cells. | |||||||||||||||||||||||||||||||||
| 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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4.6 ★★★★★
Based on 10 reviews
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Product Reviews
★★★★★ 5
The best pillow for side and back sleeping
Style: King - Medium
Bought this pillow for my husband, he loves it. He is back sleeping and has been the best pillow so far. Worth the money. With buy one for me and my daughter. It’s a little pricey but when it comes to sleeping it’s worth the money.highly recommend
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Reviewed in the United States on May 13, 2026
★★★★★ 5
Best pillow ever
Style: Standard – Low
I have to say, after decades of buying all sorts of pillows, trying to work for me and my short neck, this is THE one.
I bought the low profile ones and OMG it is just perfect for me. I'm finally getting a good nights sleep without fighting a pillow that is too firm or doubling a pillow that is too soft and thin. It's also not as heavy as other memory foam pillows I've tried. I hate a heavy pillow. Yet, it isn't so light weight that it gets pushed off the bed easily if you change positions a lot during the night.
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Reviewed in the United States on May 27, 2026
★★★★★ 5
Amazing. The Perfect Pillow for Side Sleepers with neck pain!
Size: Queen (High Loft), Number of Items: 1
I do have to update my experience. This still is a very comfortable pillow, it's just I can't sleep with it under my head (it's now my hug pillow). After using for 2 weeks, I developed severe neck spasms. Pillows are very individulized and as it turns out, I probably need a firmer pillow than this "highly" comfortable pillow can offer.
Will NOT change my rating.
Wow! I'm impressed with this pillow so far. I'm a side sleeper mostly but chose the medium-high loft 6.2" as recommended which was the perfect height. I didn't see any taller options but this has turned out just right. I did hours of research before chosing this product, which turned out to be advantageous.
It's not as cooling as my old memory foam pillow with the gel layer but it stays cool long enough that I don't have to move the pillow around to be comfortable before falling asleep. It's surprising lightweight. I like it much more than any memory foam I've ever had. The best news was that within a few days my headaches and neck pain have improved by 80%. I am really very impressed with the quality of this pillow. It is softer than I anticipated but still firm enough to keep my head elevated giving me perfect spine alignment and great support. It truly is the MOST COMFORTABLE supportive Pillow I have ever slept on!
The smell is only minor at It's worst and has nearly disappeared after just 4 days. It is a mild latex smell but definitely NOT petroleum. It's in the moderate price range at ~$100 and worth every cent. High value for the price.
I really really like this pillow and highly recommend it. Big fan! Might irder a 2nd as a backup if it doesn't last but it's so well made I feel that won't be a problem.
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Reviewed in the United States on January 22, 2026
★★★★★ 5
Almohada
Size: Standard (Mid Loft), Number of Items: 1, Size: Standard (Mid Loft), Number of Items: 1
The pillow is of very good quality—it is worth every penny spent. It is soft, and my daughter sleeps very comfortably.
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Reviewed in the United States on May 25, 2026
★★★★★ 5
Excellent comfort and support!
Size: Standard (Mid Loft), Number of Items: 1
The pillow feels premium from the first night and delivers soft and stable support. The pillow holds its shape and didn't have any chemical smell out of box. Worth the price and I would buy it again.
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Reviewed in the United States on December 30, 2025
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