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Description
Human zonulin 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. Centrifuge the specimen at 1000 × g for 15 minutes at 2-8°C within 30 minutes of collection. The supernatant can be assayed or stored at -20°C or -80°C, but avoid repeated freezing and thawing. 3. 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 the tissue and mince it. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1 g of tissue sample to 9 mL 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 the supernatant can be assayed. 4. Cell Lysis Buffer: Gently wash adherent cells with ice-cold PBS, then trypsinize and collect cells by centrifugation at 1000×g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash collected cells three times with ice-cold PBS and resuspend 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 freeze-thaw cycles or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and remove the supernatant for analysis. 5. Cell Culture Supernatant: Centrifuge at 1000×g for 20 minutes. Remove the supernatant for analysis or store at -20°C or -80°C, avoiding repeated freeze-thaw cycles. 6. 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 enzyme conjugate are sequentially added to microwells pre-coated with a zonulin capture antibody. After incubation and washing, the assay 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 zonulin in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human zonulin ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Zonulin is a protein that regulates the permeability of tight junctions between cells lining the digestive tract. It was discovered in 2000 by Alessio Fasano and his team at the University of Maryland School of Medicine. As the mammalian analog of the Zonula occludens toxin secreted by the cholera pathogen Vibrio cholerae, it is thought to be involved in the pathogenesis of celiac disease and type 1 diabetes. Gliadin, a glycoprotein found in wheat, activates zonulin signaling, independently of genetic expression of autoimmunity, leading to increased intestinal permeability to macromolecules. | |||||||||||||||||||||||||||||||||
| 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 | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.8 ★★★★★
Based on 14 reviews
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Product Reviews
★★★★★ 4
Shape can be a game changer to reduce arm/wrist pain
Style: Right-Handed, Pattern Name: Mouse, Color: Graphite
Very easy to setup with PC, whether by bluetooth, usb, wifi. It does take some time to get used to the new design. The scrolling is very good and all functions are extremely responsive. So far, it is not causing any strain to my arm and wrist which is why I am trying this design as my older, normal shaped mouse, was causing me arm pain. They should include a quick easy to understand information sheet about the buttons. Need to download in order to customize 2 other buttons. All in all, so far I like it and definitely suggest it for people having arm or wrist pain from the angle and position of a regular shaped mouse.
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Reviewed in the United States on May 11, 2026
★★★★★ 5
Wonderful!!
Style: Right-Handed, Pattern Name: Mouse, Color: Graphite
Update -
After using this mouse for over a year I started experiencing something profoundly annoying. When dragging something from one place to another (often from monitor to monitor) I would accidentally hit one of the buttons by the thumb and it would close whatever I was working on. Especially during work where I do this often. I found there is an option to turn these buttons off in the software!!! Wonderful!!!
That was the absolute only thing I was having an issue with. Longevity has been perfect and I'll confess, I'm not particularly regular about cleaning it. I would buy this mouse again in a heartbeat.
I bought this at the same time (Feb '25) as the Logitech Ergo K860 Wireless Ergonomic Keyboard - Split Keyboard, for which I have also written a review.
I cannot say enough good things about both of these products. My neck, shoulders and back feel so much better than with my previous setup.
At the time of purchase, there were a number of reviews saying these failed fairly quickly. This has not been my experience and I am not very conscientious about cleaning it.
After almost a year of heavy use (8-10 hrs a day), the batteries are at 30% which I don't think I can complain about.
These products are not cheap, but for me they have been worth every penny and I cannot see myself going back. I'd buy them again in a heartbeat.
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Reviewed in the United States on January 28, 2026
★★★★★ 5
Easy setup, beautiful red!
Color: Red, Color: Red
I needed a mouse for an assignment when I asked my son for his. I loved it so much that I needed to buy myself one. I bought the red to match my computer (and to know that the navy blue is my son's) and it's a very sleek, not tooo vibrant color. It does come with both an USB & USBC controller (see pictures) and mine connected FAST to my computer. It does NOT come with a battery (most mice won't) so please grab a DOUBLE AA battery before it comes in so you will be ready. I have no issues with the mouse and loved the price point on this one. It's light weight and easy to setup. Then again it was a plug in/play setup, so if you're not tech savvy, you won't have issues with this one. Just choose a color and get it!
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Reviewed in the United States on April 8, 2026
★★★★★ 5
Cut the Cord and Never Looked Back!
Color: Grey
I'd been using a wired mouse forever and kept telling myself the cord wasn't that annoying. Spoiler: it was. The moment I switched to the WisFox 2.4G Wireless Mouse, I realized what I'd been putting up with all along — and there's no going back now.
Setup was dead simple. Plug the little USB receiver into the laptop, and it just works. No drivers to hunt down, no pairing headaches, no nonsense. The 2.4G connection is solid and responsive — I haven't noticed any lag or signal dropout, even when the receiver is tucked into a port on the back of the laptop. The cursor tracks smoothly across my desk surface without any skipping or stuttering.
The ergonomic shape is a genuine plus. It fits naturally in the hand during long sessions and doesn't leave my wrist feeling fatigued the way some flatter mice do. The buttons have a satisfying, tactile click without being loud enough to bother anyone nearby.
My one gripe — and it's pretty minor — is that it runs on batteries, which means you'll eventually need to swap them out. It's not a dealbreaker by any stretch, and the battery life has been reasonable so far, but if you're used to a wired mouse that just runs forever, it's a small adjustment to keep a spare AA or two around.
Pros:
Instant plug-and-play setup with the USB receiver
Reliable, lag-free 2.4G connection
Comfortable ergonomic shape for extended use
Great freedom of movement without cord clutter
Cons:
Runs on batteries — plan to keep a spare on hand
If you're still tethered to a wired mouse and on the fence about making the jump, this is a solid, no-fuss way to do it. Hope this helps!
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Reviewed in the United States on May 16, 2026
★★★★★ 5
Good Work
Color: Grey
Work so good
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on June 11, 2026