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Description
Human METTL3 ELISA KitProduct Specification Usage Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High precision pipette and gun tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37 constant temperature box 4. Distilled water or deionized water Sample processing and requirements: 1. Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4 overnight, then centrifuge at 1000g for 20
Product Specification
| Usage |
Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High-precision pipette and gun tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37℃ constant temperature box 4. Distilled water or deionized water Sample processing and requirements: 1. Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4℃ overnight, then centrifuge at 1000×g for 20 minutes, and take the supernatant, or store the supernatant at -20℃ or -80℃, but avoid repeated freezing and thawing. 2. Plasma: Collect specimens using EDTA or heparin as an anticoagulant and centrifuge at 1000×g for 15 minutes at 2-8°C within 30 minutes of collection. Remove the supernatant for testing, or store at -20°C or -80°C, but avoid repeated freezing and thawing. 3. Cell culture supernatant: Centrifuge at 1000×g for 20 minutes. Remove the supernatant for testing, or store at -20°C or -80°C, but avoid repeated freezing and thawing. Pre-Test Preparation: 1. Remove the test kit from the refrigerator 10 minutes in advance and equilibrate to room temperature. 2. Prepare a gradient standard working solution: Add 1 mL of universal diluent to the lyophilized standard. Let stand for 15 minutes to completely dissolve, then gently mix (concentration 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 seven 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 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 3 (METTL3). After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by HRP peroxidase and to yellow by acid. The intensity of the color is positively correlated with the amount of methyltransferase-like protein 3 (METTL3) 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 3 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Methyltransferase-like protein 3 (METTL3) is an enzyme encoded by the METTL3 gene. This gene encodes the 70 kDa subunit of MT-A, which is part of the N6-adenosine methyltransferase enzyme. This enzyme is involved in the posttranscriptional methylation of internal adenosine residues in eukaryotic mRNA to form N6-methyladenosine (m6A). | |||||||||||||||||||||||||||||||||
| 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 | Serum, plasma, cell culture supernatant |
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4.2 ★★★★★
Based on 1084 reviews
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Product Reviews
★★★★★ 4
Worthy chew and fetch ball.
Color: L) 3" 2-Pack (Chicken)
Durable and dog loved to gnaw on it. Bright yellow color easy to keep track of for both dog and parent!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on February 16, 2026
★★★★★ 5
Flavored Dog Rubber Balls
Color: H) 2.5" 3-Pack (Beef), Color: H) 2.5" 3-Pack (Beef)
My dog absolutely loves these balls, I have a few more I had already bought, but she has an addiction to them so I had to buy more. I am able to squeeze these really easily so when she does pick them up after I’ve thrown them they’re very easy for her to pick up. They have a good thickness to them so it’s not too flimsy. It’s pretty tough and they bounce pretty high which I like and so does she know she is an aggressive chewer, but I haven’t had an issue yet with her chewing right through these, which is why I like them and we both love the fact that it smells like beef I could practically smell them through the bag, but I have a really high pitched sense of smell, and I knew she was going to love them. Not to mention you can’t beat the price.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on November 14, 2025
★★★★★ 5
Love These Balls
Color: H) 2.5" 3-Pack (Beef), Color: H) 2.5" 3-Pack (Beef)
These are great balls. My Border Collie can destroy a tennis ball in a couple of hours and the Kong brand ball is heavier than I like to play with in the house. These balls hold up to her biting and chewing but are lite weight and softer for bouncing off the walls.
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Reviewed in the United States on May 7, 2026
★★★★★ 3
Seems durable - but the scent is not the best
Color: B) 3" 2-Pack (Bacon)
I don't know if I'd call the scent of this "bacon," more like rubber with a hint of bacon. It's very strong. I gave my dog one of the balls & put one in a cabinet where I keep his treats & a few other unrelated odds & ends. I now hold my breath when I open that cabinet b/c good lord, the smell. I'm tempted to just give that one to the dog too, just because it'll air out eventually & then everything else in that cabinet won't smell bad.
My dog feels so-so about it. He'll briefly chase after it & chew on it, but loses interest in it pretty fast. My rather hyperactive cat also plays with it from time to time, as he seems to have more interest in most dog toys than the dog these days, so there's that. (Then again, he thinks a wide variety of random items around the house are loads of fun to toss about, so it's possible his opinion is to be taken with a grain of salt. Between the brief periods of one of these balls being chewed on & clawed at, it's stayed completely intact. I think if you have a dog who simply likes to play fetch or chew on rubber balls in general, this one would stand up to the task. But if you're expecting it to be greeted with the enthusiasm as a bacon grease covered ball might be - I'd adjust your expectations.
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Reviewed in the United States on February 19, 2023
★★★★★ 5
My dogs favorite toy so far in two years
Color: C) 4" 2-Pack (Bacon)
I use this with my medium sized dog. These are large for him but that's what I want so I don't ever have to think about them being too small or a swallow hazard. He has been through A LOT OF TOYS in the last couple years, but nothing, NOTHING has he loved as much as these.
His favorite used to be squeaker balls. Then his favorite was crackle balls. But he will gladly ignore both for this one. I don't know if it's the bacon flavor or what, but he go nuts for it.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 5, 2026
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