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Description
Rat AQP5 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 ultrasonically disrupted or repeatedly frozen and thawed. Finally, the homogenate is centrifuged at 5000×g for 5-10 minutes and the supernatant is collected 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 utilizes 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 an aquaporin 5 (AQP5) capture antibody. 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 aquaporin 5 (AQP5) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Aquaporin 5 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Aquaporin-5 is a protein encoded by the AQP5 gene. AQP5 is a water channel protein. Aquaporins are a family of small integral membrane proteins related to the major intrinsic protein (MIP or AQP0). Aquaporin-5 plays a role in the production of saliva, tears, and pulmonary secretions. AQP0, AQP2, AQP5, and AQP6 are closely related and all map to 12q13. | |||||||||||||||||||||||||||||||||
| 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 | Tissue homogenates and other biological fluids |
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4.6 ★★★★★
Based on 28 reviews
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Product Reviews
★★★★★ 4
Accurate and Durable – A Game-Changer for Long Cooks!
Size: 2 Probes
After a lot of research, I decided to invest in this wireless meat thermometer, and I’m really glad I did. My first cook was a brisket, and it helped me keep an eye on the temperature with ease. The durability of these thermometers is impressive – they feel solid and built to last, even with regular use around the heat and elements of a smoker.
In terms of accuracy, they’re spot on. The temperature readings have been reliable, which is essential for any serious BBQ enthusiast. These thermometers take the guesswork out of smoking, and they really do work well.
The only downside I’ve noticed is occasional connectivity issues. A few times, the connection dropped and it stopped reading, which I’m still troubleshooting. I’m not sure if it’s due to the distance, as the reader is usually inside the house, or if there’s another factor at play. However, I learned that for long smokes, it’s smart to alternate between the two thermometers to keep them charged and ready for the duration.
Overall, these thermometers have been top tier, making smoking and grilling much easier and more precise. If you’re willing to keep an eye on the connection, this is an excellent addition to any BBQ setup.
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Reviewed in the United States on November 3, 2024
★★★★★ 5
A must-have high-tech gear for cooking
Size: 2 Probes, Size: 2 Probes
I've been looking at wireless thermometers for while but was unsure about which brand to choose. Then I saw someone on YouTube use the Sync to cook a Christmas turkey and decided to give it a shot. The youtuber didn't just cook the entire turkey as a whole. Instead he cut it into different parts, and cooked them simultaneously at two different durations for optimal results. This was ingenious as dual probes can be extremely helpful for certain foods that require different doneness, and this thermometer monitors both the internal temperature of the food and the ambient temperature.
Fast forward two weeks in, and I'm in love with this thermometer. Even though I have an instant food thermometer, I still find myself using the Sync more because you can leave the probe in to get readings instead of having to manually stick the probes in like you would with a traditional food thermometer. As an experienced chef, even though most foods look cooked on the outside, you can never really judge their internal doneness without a thermometer.
1. Highly accurate: Each probe has 5 sensors for multi-zone temperature readings ensuring the lowest point of the internal temperature is displayed with accuracy down to the decimal. I cowitnessed the readings with a high-end instant-read thermometer, and all the readings were consistent with each other. It also comes with an external ambient temperature sensor which is important because, ambient temps may vary in different parts of the oven/smoker/grill. I cowitnessed the ambients temps as well and they were also consistent.
2. Easy to connect: It can easily connect to my phone via wifi or Bluetooth. Once set up, it's super easy to use every time.
3. Stable connection: The wifi or Bluetooth connection is very stable. I've used Sync in a smoker for several cooks now, and even when it was wrapped in foil with a the lid of the smoker on, it never lost connection. There were no complex syncing issues or signal losses.
4. You can monitor the cooking progress from anywhere in your home where you can take your phone or tablet. You don't have to stay in the kitchen.
5. You can set preferences for various alerts via the app.
6. The built-in cooking time estimator is great when you're in charge of larger meals or just want to know when to lower the heat. The app will tell you when the food is done and allow a rest period for the temperature to rise. For example, if you want a medium steak and set the temperature to 135℉, the app will tell you the steak is done when it reaches 130℉ and to allow a 3-5 minute rest period for the temperature to rise to 135℉. This means you can cook proteins perfectly every time.
7. It has a large base screen that's easy to read.
8. Charging is very convenient. Just plug in the charging cable to charge the base, and stick the probes into the base to charge the probes.
9. You can select a preset in the app modify it according to your preferences.
10. Customer service. Initially, when my Sync was not charging, I contacted customer service. To my surprise, they responded quickly and offered to replace the device for free if it was defective. I was very impressed with their quick response and friendly attitude.
11. The 3-year warranty is a nice bonus.
Hopefully this review has been helpful. I'll update this review if my views change or I find more ways to use it. If you're concerned about the price being higher than entry-level products, then you probably haven't taken the time to read this.
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Reviewed in the United States on February 15, 2024
★★★★★ 5
Efficient, accurate, and thoughtfully designed
This instant-read thermometer from Typhur does exactly what it promises, quick, accurate readings with no fuss. It’s fast, reliable, and has become one of those tools I reach for constantly in the kitchen.
The display is bright, clear, and easy to read. It even auto-rotates based on how you're holding it, so the numbers are always right side up, no more twisting your neck trying to read upside-down temps.
One of my favorite features is the magnetic back. Being able to attach it to the fridge keeps it visible and within arm’s reach when I need it most, no digging through drawers mid-cook. It’s a small detail, but one that adds a lot of everyday convenience.
Unlike some of Typhur’s other products that charge via USB-C, this one runs on batteries. At first I found that a little unusual, but on second thought, I actually appreciate it. No internal battery means no long-term degradation, just pop in fresh batteries when needed and keep it going. That kind of longevity makes sense for a kitchen tool like this.
Best part? I scored it on sale for under $40, which was a steal. Even at its current price point around $100, it still feels fair for the quality and performance you're getting.
Highly recommended if you’re looking for an instant-read thermometer that blends speed, accuracy, and solid design.
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Reviewed in the United States on June 4, 2025
★★★★★ 5
Precision meets style - a game changer for culinary enthusiasts
Just received Typhur InstaProbe my latest kitchen gadget, an ultra-premium meat thermometer, and let me tell you - this is not your average thermometer. It cost about a lot more than the standard meat thermometer, but every single penny is worth it for the level of accuracy and convenience it brings to the table.
The speed and precision of this thermometer are unrivaled. It takes readings in a split second, completely eliminating the need to wait around while your hand hovers over the heat, or loss of precious heat from your oven. The accuracy is pinpoint - to the decimal. No more guesswork, no more undercooked or overcooked meats. Every meal I've made since getting this thermometer has been cooked to absolute perfection, making me feel like a gourmet chef in my own kitchen.
The aesthetic and tactile quality of this thermometer is another feature that sets it apart. It's sleek, modern, and feels premium in your hand. It's not just a tool, but an accessory to my kitchen that is both functional and pleasing to the eye.
Despite the price tag, this thermometer is worth considering for those who value precision, speed, and style in their cooking. The level of control it gives you over your cooking is unparalleled, and it's incredibly easy to use. It's a must-have for serious home cooks, barbecue enthusiasts, or anyone who values precision in their cooking. It's a worthwhile investment that will transform your cooking experience and elevate your meals to new heights. And as the old saying goes - you get what you pay for. With this premium meat thermometer, you're getting the very best.
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Reviewed in the United States on June 12, 2023
★★★★★ 5
Great quality
I’m just a hobbyist, but this is the best fast read temperature probe I’ve ever used. Performance is top notch. The build quality is excellent, especially the display screen. You will be able to read it in any light situation. The temperature reading stabilized very quickly, ambidextrous response is also very fast. Obviously to turn the unit on you will need to swing the probe out. It has good magnet strength, but, in cases where you have a true stainless steel grill or prep table, just lay it down, in out position, it will awaken within a second when picked up. Close the probe, it will stay off till next use. The box it is shipped in makes for great storage, just make sure to keep the outer sleeve of the box. The probe itself is very sharp (but, be careful) unlike some others, which makes it very easy to push in to the most delicate, to the very tough cuts of meat at any temperature. It uses 2AAA batteries which are are very versatile, unlike the 2032 lithium batteries that others use. I understand that alot of people use these products in cold temperatures in which alkaline batteries will not work well, but you can get AAA lithium batteries for cold weather conditions. Just make sure you remove the alkaline batteries when the probe will not be used for long periods of time due to corrosion. All in all, a great product, it’s built with quality materials and should last a very long time. Quality over quantity. Also, they offer a 10 year warranty once you register your probe. They actually reached out to me after I purchased it and left a phone number. I called them and actually spoke to a real representative of the company. You may pay more for this temp probe than 100’s of others that are less expensive, but, I feel that Typhur stands behind their products.
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Reviewed in the United States on June 26, 2024
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