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Description
Human MGAT3 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 the specimen using EDTA or heparin as an anticoagulant and centrifuge it at 1000×g for 15 minutes at 2-8°C within 30 minutes of collection. The supernatant can be tested or stored at -20°C or -80°C. However, repeated freezing and thawing should be avoided. 3. Urine, saliva, sweatOther 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 beta-1,4-mannosyl-glycoprotein 4-beta-N-acetylglucosaminyltransferase (MGAT3). 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 beta-1,4-mannosyl-glycoprotein 4-beta-N-acetylglucosaminyltransferase (MGAT3) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Beta-1,4-mannosyl-glycoprotein 4-beta-N-acetylglucosaminyltransferase ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | β-1,4-Mannosyl-glycoprotein 4-β-N-acetylglucosaminyltransferase is an enzyme encoded by the MGAT3 gene. Over 100 different glycosyltransferases are believed to be involved in the synthesis of protein-bound and lipid-bound oligosaccharides. The enzyme encoded by this gene transfers a GlcNAc residue to the β-linked mannose of the melamine core of an N-linked oligosaccharide, generating a double-stranded GlcNAc residue. Multiple alternatively spliced variants encoding the same protein have been identified. | |||||||||||||||||||||||||||||||||
| 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 | 0.15-10 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, urine, saliva, sweat, and other biological fluids |
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4.9 ★★★★★
Based on 14 reviews
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Product Reviews
★★★★★ 4
Fantasy
Format: Hardcover
Not my favorite genre however the book is written really well and my students who love fantasy loved this book
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Reviewed in the United States on November 20, 2025
★★★★★ 5
Brilliant
Format: Hardcover
The First State of Being is a brilliant, highly readable middle grade book from the QUEEN of character-driven middle grade literature. This expansive sci-fi feels at once introspective and cinematic, leaping off the page like something made for a movie. This book will help young people and all readers reflect on our past and future as a human community, especially in terms of health advances, animal extinction, and the potential for technological development. It also highlights the love and fight in a mother and the need to view life through a positive lens by focusing most on the present, not our past or future.
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Reviewed in the United States on February 2, 2025
★★★★★ 5
Which Came First?
Format: Hardcover
Of course this won the Newberry award. From the first word, it reminded me of Donna Barba Higuera's The Last Cuentista. A plot so different and imaginative that you wonder how anyone could have thought of it and then carried it out. It's a story of a young boy who is trying to take care of the single mother who seems unable to take care of the family, through no fault of her own. It's a sci-fi fantasy. And it's a story of taking care of each other. At the totally satisfying end I found myself wondering about the chicken and the egg. Thinking I may have to read this one again to see if I can figure it out.
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Reviewed in the United States on March 3, 2025
★★★★★ 5
Perfect Choice
Format: Hardcover
Excellent YA / SCI-FI novel. Pretty well captures the spirit of a 12-year-old boy.
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Reviewed in the United States on December 25, 2025
★★★★★ 4
Well-written, twisty novel for middle school children
Format: Kindle
This is Kelly’s second Newbery Medal, the first being for *Hello, Universe* in 2018. Both are well-written novels for middle-grade children and should have a lot of appeal for that age.
It's the year 1999, early fall, an apartment complex in Delaware. 12-year-old Michael is worried that the Y2K bug will cause the collapse of civilization on January 1, 2000. He has been stealing cans of food to create an emergency stash under his bed. He has started a new school, and his only friends are the apartment complex maintenance man and his high-school-age after-school babysitter, Gibby. Michael’s father is dead, and his mother works long hours to make ends meet.
One day a strange kid shows up in the complex, dressed in unusual clothing and asking odd questions like, “What year is this?” It doesn’t take an experienced science fiction reader to suspect that this kid, Ridge, is from the future. He is interested in observing late 20th Century American culture. But he also wants Michael and Gibby to help him figure out how to get back to the future without interfering with the past and thus changing his own timeline. The 1999 part of the story is interspersed with notes about the future and the panicked dialog of Ridge’s family as they try to investigate what happened to him.
It probably doesn’t have many new ideas for adult readers, but it will be surprising and exciting for readers ages 12 and up
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Reviewed in the United States on September 9, 2025