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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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4.5 ★★★★★
Based on 7 reviews
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Product Reviews
★★★★★ 4
Tough STINKY ball
Size: Large, Color: Red
This ball is the only one I've found that my dog can not destroy in seconds. He can chew on it for a few weeks before he works a hole in it. That's the good part. The bad side is the horrible smell! It doesn't seem to bother him, but it is horrible to my nose. It is so awful that I tried soaking it and dish soap and vinegar water overnight to see if I could make it go away. It was a little bit better but still stunk. Anybody got other suggestions on what could make the smell go away?
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Reviewed in the United States on April 12, 2026
★★★★★ 5
A good toy for chewers!!
Size: Large, Color: Blue
My dog loves this ball! The best part is that he hasn't destroyed it yet. He's a real chewer and has destroyed several toys since I got him about 6 months ago. The ball is very solid and a the squeak is not annoying. A great toy
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Reviewed in the United States on May 28, 2026
★★★★★ 5
My dog loves it, holds up well
Size: Large, Color: Blue
My German Shepherd absolutely loves this ball! He is a pretty aggressive chewer and it has held up really well, not indestructible but it lasts. We currently have three and he always has one in his mouth.
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Reviewed in the United States on May 8, 2026
★★★★★ 5
Lasted a year with a Great Dane
Size: Large, Color: Red, Size: Large, Color: Red
It lasted an entire year as my Great Dane’s favorite squeaky ball. The only reason it’s failing is because the squeaker no longer works great.
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Reviewed in the United States on May 31, 2026
★★★★★ 3
Not indestructible but pretty darn tuff for sure!
Size: Large, Color: Blue
I would have maybe given a better rating if they hadn’t lied about it being indestructible and recommended it for my dogs size range! It didn’t last the first day. However my dog may be a bit above average when it comes to his relationship with the balls! It’s an issue and he should probably be in a 12 step program of some kind. But we all have our vices right and his is balls. All of them he needs em and he never stops joining for them and the stronger they are the more he’s driven to defeat it! So now that you have full disclosure from me. The ball is still in his box and it’s still round and mostly intact and that’s after several months! The part that didn’t last the first day was the squeaker! Poor little dude! If he would have just stayed quite it. But he didn’t and Bo dog knew he was in there! And made short work of him. Other than that. The ball pretty dang tough! But it’s not indestructible is it a big deal. Well in this world of way to many choices and then having to find the few that will tell you honestly what your getting and are not over stating their product! Just to get your money. And couldn’t care less about the quality! It sucks that we have to navigate all the crap to find the few decent things available. So it most certainly does matter! And I say that for their benefit the most hopefully! Because people are craving and constantly looking for a company that just makes the effort to put out the best they can. And tell you the honest description of what they have. That stands behind their work and puts more effort into quality then they do into profit. With the faith that the quality and integrity in their company will end up bring them the profits they can be proud of and know that they earned! Honestly! Is this company one of those! I don’t know? It’s not clear and in their defense I never reported the sneaky man’s demise on day one! I expected that. But indestructible is a descriptive word that has a very specific meaning. And when it’s used in a manner that it was! Then you’ve set yourself up to fail as there are very few things that meet that description! And I know I’m picking hairs here. But it’s words that get used like that wich makes one appear to have not met the standard they wanted to! And maybe you don’t realize its impact on your overall until it’s far down the road! Can it be said better. Sure . Always room for improvement. But I would certainly remove the claim that’s just unreasonable to expect. If I wanted to use the benifit the does have to offer perhaps I would have said this ball is the closest thing to indestructible we’ve made so far. That’s statement tell me that they are not claiming it is but that it’s pretty tough! And it with out exact words tells you these guys must take honesty seriously or they wouldn’t have chosen to claim the accurate step below. And that lets me know that they hold themselves to a standard that they obviously respect their customers and they don’t take them for granted that’s integrity right there and there’s just not enough of it o. This world today! Seems to me that there are a few that are really close to it though. Maybe so close it’s worth the time to just go ahead and say it. If my gut feeling is right then it’s absolutely worth the time! Do I recommend the item absolutely it’s as tough as nails! And if you hate the squeaky part. Then don’t worry it won’t last long if your dogs like mine. Or you can take the little dude out when the dogs sleeping! And I’m going to add a star because when I look at it in the wider view I’ve been a bit harder then is justified I believe. Their. Our lives today are not as good as they once were and should be! Because of our bad habit of not holding ourselves to the standards we are capable of and rather settling for what’s out there at the time. Because of many reasons right. What’s the point. Or it’s not going to change a thing. Or that as good as it needs to be. Don’t want to seem so picky and all the justification we inject in the place of our potential. That’s had a vey negative effect across the board! And we’re losing so much more then quality dog toys as a result. When we let or standards drop due to whatever reasons as a civilization then are standards drop. And that’s all you should need to know. To understand where that leads! And we only do ourselves the disservice and loss that comes from those choices. So look around and don’t hesitate to. All out that when you see it. Just try and come from a place that’s not to tear things down but to remind them of what they are really capable of! And how much we all need to hold ourselves to the highest standard we are capable of! And stop justifying excepting less! And then we see the results from that rather than the other.
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Reviewed in the United States on August 18, 2025