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Description
Human METTL16 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 capture antibody against Methyltransferase-Like Protein 16 (METTL16). 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 Methyltransferase-Like Protein 16 (METTL16) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Methyltransferase Like Protein 16 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Methyltransferase-like protein 16, also known as METTL16, is encoded by the METTL16 gene. It directly binds to U6 snRNA in vivo. It associates with various ncRNAs, lncRNAs, and pre-mRNAs. Its N-terminal module is essential for charge-based RNA binding. It preferentially methylates adenosines in structured RNAs. It recognizes the sequence and structure of methylated substrate RNAs. The methylation activity of METTL16 can be regulated by the conformation of the substrate RNA. It is autoregulated through its SAM binding pocket. Mutations in its catalytic domain modulate MAT2A mRNA levels in cells. Its regulation of MAT2A mRNA is crucial for embryonic development. | |||||||||||||||||||||||||||||||||
| 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
Received giftcard money to replace my plan-protected malfunctioning motherboard
I've been subscribing to the Asurion protection plan for a while. Recently, an Asus Prime PC motherboard I had bought a few months ago on Amazon stopped working well. I ended up considering filing a claim with Asurion to see if they would either fix/replace it or issue the purchase price money, so I could buy the failed electronics replacement component again for my custom-built PC. I went to the protection plan's website and started the claim filing process but couldn't find the category my failed component would be under in order to select it from the drop-down list. I ended up messaging the live customer service. The chat messaging was somewhat slow, the agent told me they were experiencing some communication delays throughout their systems. Ultimately, the agent was very kind and understanding, and completed the claim filing process for me on her side by asking me to provide the necessary info such as the item's product name, date of purchase, price before taxes, etc. I received a printable UPS shipping label in the email, printed it, and returned the item in the mail. Approximately 2 days later, I received an Amazon giftcard code in the email with the dollar amount equal to the item's price before the sales tax. Then I redeemed the giftcard code on the Amazon page and proceeded to apply the giftcard money to my next purchase of a replacement Asus prime motherboard, and it all worked out successfully. Therefore, I had no issues with the protection plan service and recommend it to others.
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Reviewed in the United States on May 24, 2026
★★★★★ 4
Fast and easy but I dont trust the process to be fully automated.
The process was very smooth and fairly quick. The only challenge I had was that the CLAIM status showed complete but I had to manually go look up the status. There was no email or notification it was closed. After manually checking the status, I called customer support and they said since I called in, they would process the payout. I wonder if I didnt call in if they would have processed it? Also, the phone rep said they would issue me a amazon egift card but then the next day I received a email saying they mailed out the payout. Very poor communication but so far the process was easy and fast.
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Reviewed in the United States on April 1, 2026
★★★★★ 5
My great experience with Asurion
Very easy to use, no hassle, everything was done quickly online (5 minutes or so), and the reimbursement check was received promptly. Exactly as advertised. Make sure you keep your Asurion info in a folder that you can easily find in a couple of years. I guess it matters on what you are protecting, but for me and my gaming chair, this was so worth it, worth it, worth it!
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Reviewed in the United States on November 24, 2025
★★★★★ 5
ASURION REFUND
I couldn’t be more pleased with ASURION. I had a 3 year furniture protection plan, when I emailed them with my problem they did not hesitate. I got a response from them quickly. It is well worth the money to get a Protection Plan.
I received a check from ASURION for the price I had paid.
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Reviewed in the United States on May 11, 2026
★★★★★ 5
A must have
I thought this was going to be a waste of money but I couldn't have been more wrong. I purchased this for a piece of furniture and after about two and a half years the piece started falling apart. So again I thought I was wasting my time but I called them up and the person that answered the phone was the kindest and the most understanding person. I proceeded to tell her why I was calling and of the issues I was having and within just a couple minutes and just a couple more questions I had an Amazon gift card for the amount of the full purchase price, I received a full refund. And this was after two and a half years. The bottom line is you can not go wrong with the Asurion 3 year Protection Plan. A MUST HAVE!!!
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Reviewed in the United States on February 10, 2025