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Description
Human FUT2 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 galactoside 2-alpha-L-fucosyltransferase 2 (FUT2). After incubation and washing, the sample is developed using the substrate TMB. TMB converts to blue under the catalysis of HRP and to yellow under the action of acid. The intensity of the color is positively correlated with the amount of galactoside 2-alpha-L-fucosyltransferase 2 (FUT2) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Galactoside 2-alpha-L-fucosyltransferase 2 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Fucosyltransferase 2, also known as FUT2, is an enzyme encoded by the FUT2 gene. FUT2 is a phospholipid peroxidase that protects cells from membrane lipid peroxidation. The antioxidant enzyme fucosyltransferase 2 (FUT2) belongs to the glutathione peroxidase family, which consists of eight known mammalian isoenzymes (GPX1-8). FUT2 catalyzes the reduction of hydrogen peroxide, organic hydroperoxides, and lipid peroxides at the expense of reduced glutathione and plays a role in protecting cells from oxidative stress. | |||||||||||||||||||||||||||||||||
| 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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4.3 ★★★★★
Based on 653 reviews
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Product Reviews
★★★★★ 4
True history
Format: Hardcover
Good information
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 21, 2026
★★★★★ 5
Outstanding book!
Format: Hardcover
A great read!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 1, 2026
★★★★★ 5
... this book was instrumental in planning and is highly recommended. In a somewhat bullet format I'll add a ...
Format: Paperback
We finished the Wonderland yesterday- this book was instrumental in planning and is highly recommended. In a somewhat bullet format I'll add a few notes...
I think it is better to deliver your food caches rather than mail them- discuss with the rangers THE ACTUAL PICK-UP POINTS, and have your map with you- they will give you current trail and water conditions, this proved to be very valuable.
We opted for a 12 day hike, after just a few days we realized that we had underestimated our hiking ability and changed out schedule to a 9 day hike. The rangers were very good about helping us with the changes, keep a slower plan and a faster plan in mind as the hike progresses (don't forget to get word to the rangers to re-date your food caches if you change your schedule).
Thank those rangers and back country guys when you see them-the trail is in splendid condition, the maintenance is ongoing. When you meet a man wearing a pack-board with a 55 gallon drum lashed to it and he is going to muck out one of the outhouses on the trail is really makes you think about all the steps that you are traversing, the water bars, the brush...on and on with the list.
Trail-trash... and I'm not talking about litter, there are some not so nice people on the trail who think nothing about leapfrogging a campsite because they "didn't like it" or the day was too long / short. That means that if you are a late arrival at a camp all the sites may be taken, then you have to ask to see permits and engage in eviction, the rangers need to hear about these guys and they leave the park "With extra paperwork:". One party we met started as a party of 3, but one of them became ill and suffered a fall resulting in an injury- so she was abandoned at Golden Lakes to fend for herself... dumped on the rangers. Point being, know your trail team- when you look at the map note where the roads are close by the trail should you need to get help.
The authors mention bugs... perhaps once or twice... they talk about deer flies and horse flies... but as much as they talk about it is still under emphasized, there are legions, hordes, armies of bugs at some of the campsites DON'T IGNORE THESE WARNINGS.
There was a great tip about putting apples / oranges / pears in your food caches- after a couple of days without fresh fruit these were a huge bonus. We also included some of the Hormel dinners- already hydrated and a nice break from the normal trail food.
All in all, this is essential equipment in planning your hike.
The Wonderland is aptly named- this one of the high churches in the outdoor religion, you will be constantly amazed!!
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Reviewed in the United States on July 16, 2015
★★★★★ 5
The Best Wonderland Trail book out there!
Format: Paperback
To plan my Wonderland Trail hike, I bought three different books, including this one. Tami's is by FAR the best. It offers great summaries of different sections as well as detailed descriptions. So many details are covered: permits, the Spray Park and Northern Loop options, packing, caching, conditioning, etc. There are numerous maps, and all of the charts at the back really helped me plan a well-informed itinerary. Great book!
I used a hard copy edition of this book to plan my Wonderland hike (since I still really like good old-fashioned books) and then downloaded the Kindle version to my phone to use for reference on the trail. After a quick read each night, I could hit the trail the next morning well-prepared for the day ahead!
I should point out that I did also bring along a Green Trails map of the Wonderland Trail, which I really only wanted to identify mountains, etc.
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Reviewed in the United States on September 6, 2017
★★★★★ 5
Best guide book I've ever purchased
Format: Paperback
This book is outstanding! Even though we backpacked the Northern Loop instead of Wonderland, there were sections in the book covering some of the camps we hiked to over our five nights in the back country. And then I ran the Wonderland Trail over three days the following week, so I was eagerly reading each night in my tent to see what I would be discovering on the trail. The author is a very good writer, knows her craft extremely well, and has obviously spent ample time in this absolute treasure of a national park. Thank you so much for writing such a valuable book.
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Reviewed in the United States on August 26, 2020