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Description
Human AQP-3 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 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, biotinylated detection antibody, and HRP conjugate are sequentially added to microwells pre-coated with an Aquaporin 3, Gill Blood Group (AQP-3) capture antibody. 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 Aquaporin 3, Gill Blood Group (AQP-3) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Aquaporin 3, Gill Blood Group ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Aquaporin-3, the protein product of the AQP3 gene, is present on the basal cell membrane of the primary collecting duct cells, providing a pathway for water to leave these cells. Aquaporin-3 is also permeable to glycerol, ammonia, urea, and hydrogen peroxide. It is expressed in various tissues, including the skin, respiratory tract, and kidneys, as well as in various types of cancer. In the kidney, aquaporin-3, unlike aquaporin-2, does not respond to vasopressin (also known as antidiuretic hormone). | |||||||||||||||||||||||||||||||||
| 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 | 0.31-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.6 ★★★★★
Based on 2369 reviews
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Product Reviews
★★★★★ 5
Inexpensive doesn’t mean cheap.
Color: Black/Orange
After 10 months, zero complaints. Well, I got rid of the horrible NATO strap 🙄. That’s a personal preference. Easy to swap out to whatever you like. Keeps accurate time meaning it’s the same time. I’m not micro analyzing it and the seconds gained/lost. It has a confident look and feel and makes you feel good when wearing it. Sure it’s not high priced but it’s not cheap. Very comfortable, light and not unincumbered. If this is what you’re looking at, you’d be satisfied. Great looking, durable and simple watch. Full confidence.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on October 25, 2025
★★★★★ 3
Budget Seiko Prospex this is not
Color: Black/Orange
Bought this as a cheap work watch, but I guess all the budget went into the solar technology.
No date adjust feature, gotta crank the crown over and over to adjust the date. (Also no screw-down crown, but that's expected at this price)
Fake diver bezel. Lol why? Cringe as heck. Should of just put a compass bezel on there instead tbh.
Cheap plastic crystal, scratches and mars just by looking at it.
Okay NATO strap. Falls apart quickly, but it's easily replaceable with a better quality one.
Decent lume on the hands, but none on the hour markers even though it looks like there is.
Looks cool.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on June 13, 2025
★★★★★ 5
Simplicity
Color: Black/Orange
This watch was received exactly as advertised. First off the watch is economical compared to other watches that advertise the same details. I will say that I have gone through numerous Luminox watches only to be disappointed. This watch fits the bill. It does have solar cells which mean it needs to be exposed to light to keep running, not a problem. Three minutes under my surefire light I am golden. The bezel doesn't rotate actually no big deal for me. I have always required that the bezel moves not anymore after having a bezel that rotates I use it next to never. Here's why, the more I rotate the bezel the more the numbers wear off. The strap on the watch with the catches for the strap are perfect. They actually hold the watch strap in place in lieu of like other big name brands that just flop and slide. As far as everyday use goes this watch is my go to. To be honest I have purchased three. This is the only watch I will wear moving forward, yeah the glass my scratch but even the big names like the Luminox I spoke of earlier scratches too.
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Reviewed in the United States on April 4, 2026
★★★★★ 5
So far so good
Color: Brown
I have had issues with other models/variations of the timex expedition in the past, so I was hesitant to get an even more expensive version of it, but I just really liked this watch so I gave it a shot. I was looking for a new “daily driver” watch as I have a bunch of nicer, more expensive watches that I didn’t want to wear to work at a hospital. I like the solar charging feature for sure as I change my own watch batteries and it can be a pain sometimes. It’s a nice weighty watch, which I like because it feels solid, not cheap. So far it seems to be keeping time well, which is an issue I’ve had with other expeditions in the past. I really like the look of this watch a lot. It’s a sharp looking watch without being too flashy or fancy so it can be worn with just about any attire. I will note that because of the solar feature, it needs to be charged well before setting the time. I placed it under a led light for about 24 hours before setting the time and using it and when I don’t wear it I place it near a window. So far I’m really liking it, but I haven’t had it long enough to really know about long-term reliability. I will update if any issues arise.
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Reviewed in the United States on September 14, 2025
★★★★★ 5
Timex Expedition Solar a no frills fine watch
Color: Brown/Blue
The watch came uncharged. I guess you're supposed to charge the watch by exposing it to sunlight. There was no sunlight and I wasn't about to wait for any. I placed it in front of one of my ultra-bright led flashlights for about 5 hours that night. It's been working fine ever since. I'll expose it to sunlight when I see some. It's a nice looking watch, no bells and whistles but it's been keeping excellent time. I swapped out the band for a NATO which I find more dependable. The price shouldn't put you in the poor house and if it's like all the other Timex watches I've owned, I'll be happy with it.
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Reviewed in the United States on April 5, 2026
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