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Description
Rat AGTR2 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 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 angiotensin II Receptor 2 (AGTR2) 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 angiotensin II Receptor 2 (AGTR2) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Angiotensin II Receptor 2 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | The angiotensin II receptor 2 (AT2R), also known as the AT₂ receptor, is a protein encoded by the AT2R gene. This gene encodes AT2R, an integral membrane protein that binds to two different G protein-coupled receptors. This receptor functions to mediate signaling in pulmonary fibrosis and regulates nitric oxide synthase expression in the pulmonary endothelium. AT2R has recently been identified as a target for the treatment of lung inflammation in cystic fibrosis (CF). | |||||||||||||||||||||||||||||||||
| 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 | Tissue homogenates, cell lysates, and other biological fluids |
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4.6 ★★★★★
Based on 7 reviews
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Product Reviews
★★★★★ 5
Great Product. Easy to use.
Color: Black
This is the third running watch I've had and it's my first Timex. I've owned to Casio's before (one being a g-shock), but after finding my g-shock to bulky and cluttered, and loosing my other Casio I picked up from Walmart, I decided to look around to find a new durable running watch.
I run cross country and track so going from a generic stopwatch to a 30-lap chrono watch has been sweet. The lap timer is easy to use and the display is big enough to take a glance and see my split and total time side by side. The recall feature works perfectly and you even have some room to adjust the settings to configure your lap display how you like it.
My favorite thing about this Timex, is its ease of use. The only time I've had to look at a manual is when I first got the watch. The display updates as you go through the various modes to show you what the new buttons mean so there is never a question on how to set the time or recall your laps.
This watch is extremely durable. It has already taken a few shots and I've swam with it on several times. The screen and body have held up nicely and I don't foresee any problems with them in the future. The strap is great also. I really like the notch that keeps the excess strap in place.
The countdown timer and the alarm both work as expected. The timer has some different settings and is easy to set up. My only disappointment with the timer was that you can't set different intervals. Like if you want to run for five minutes and walk for two your going to have to invest a little more for a watch that allows different intervals. The alarm works well too, and the beeping successfully wakes me up in the morning, but I'm not a heavy sleeper so some may find it too quiet.
Overall this watch is durable, easy to use, and can track splits really well. The watch is a little bulky if you are used to a more minimalist watch but for me the weight was perfect, and for the price you can't go wrong with this watch.
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Reviewed in the United States on June 5, 2013
★★★★★ 4
Trusted Brand Delivers Again
Color: Black/Yellow
I had my previous Timex watch for 30+ years and it finally died after getting wet, so I replaced it with the closest model I could find to it.
PROS:
- Has the basic functions I needed: Clock, 2nd Time Zone Clock, IndiGlo backlighting, Alarm, Timer, Stopwatch. Allows me to leave phone at home and just take this out.
- Easy to set time/date/etc. Can turn alarm on / off easily. Buttons for timer/mode switching work fine.
- Clear display so easy to read. The backlighting makes this even better.
- Can swap out the band (I put mine on a Chums' "The Band" that I like) if the new one takes 19mm pins.
CONS:
- The only thing I dislike is it is heavier than my old Timex. A little bulkier too. Not annoyingly so but noticeable enough for me to comment on it here.
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Reviewed in the United States on October 20, 2025
★★★★★ 5
Great Timex Watch
Color: Black/Yellow
This Timex Ironman watch arrived on time and is of great quality. I have used these watches for years and have NO complaints about their longevity, accuracy, or dependability. I will definitely purchase again if needed.
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Reviewed in the United States on May 1, 2026
★★★★★ 5
Item as described.
Color: Dark Blue
A timeless tradition of a great design and useful watch. Have been using this watch design for over a 30 years.
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Reviewed in the United States on May 27, 2026
★★★★★ 5
Great thowback to the OG Ironman, but Amazon's listing gets it undue negativity.
Color: Black/Yellow
Amazon's listing is not very good with it's wording so this watch has gotten undue negative reviews.
I've seen this model listed as both the Endure 30 and the Original 30 Shock, it has 1 alarm with 3 different modes (not 3 separate alarms), 200m WR, ISO shock resistance, (reverse) Indiglo with night mode, 2 time zones, 12/24hr time, 30 lap stopwatch, 24hr countdown timer (repeatable), and day/date (MM.DD or DD.MM). Its basically a slightly updated feature set compared to an Ironman 8-lap.
This watch is great, it's got the look of the original Ironman 8-lap with modern guts. The only minus for me is it could be a little slimmer on the wrist, but I also didn't realize it was shock resistant when I bought it. For comparison, it is a few mm smaller in all dimensions than a G-Shock G2300/G2310/GW2310 series. The band is similar to G-Shocks in that it is formed/molded around the wrist but like the case it's still slimmer in the way it wears around the wrist. Not as slim as an F91W but not as massive as any G-Shock basically. The module has a better display with bigger numbers than the above mentioned Casios. With the exception of the lap memory, the G23## G-Shocks have more features, but the Endure 30 is much easier to use thanks to the display and larger buttons.
If you want 3 alarms you need the very similar Classic 30. The main differences being the Classic has 3 separate alarms (not 1), occasion reminders and 3 time zones but losses the Ironman 8-lap look, the shock resistance and it's only 100m WR. The Classic seems to come in at least two case varieties (chunky or slim), two sizes and many color combinations.
If you only need the Endure 30's features but want a different shape/size/style/slimmness, I believe the Essential 10/30 and BASIC Transit models are functionally the same with only different lap memories, WR, and no shock resistance.
Unfortunately, Timex doesn't easily identify the actual module used in a watch like Casio, so the best way to figure out what features a watch has is to lookup the watch model on the Timex website. Of course the manuals do not always match the marketing names they have used over the years (Endure/Classic/Essential/etc), and each manual covers a few shapes/sizes of watch but just search for the model number in the manuals sections and you'll eventually find the right one. If no manual pops up right away delete digits from the right end of the model number until a manual is found, I believe those last digits only indicate slight variances in style/color that are not important to functionality.
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Reviewed in the United States on May 31, 2021