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Description
Mouse OXTR 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 an oxytocin receptor (OXTR) 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 oxytocin receptor (OXTR) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Oxytocin Receptor ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | The oxytocin receptor (OXTR) is a protein that functions as a receptor for the hormone and neurotransmitter oxytocin. The OXTR protein belongs to the G protein-coupled receptor family, specifically Gq, and acts as a receptor for oxytocin. Its activity is mediated by G proteins, activating several different second messenger systems. It is expressed by mammary myoepithelial cells and in the myometrium and endometrium during late pregnancy. It is also associated with the central nervous system. The gene is thought to play an important role in social, cognitive, and emotional behavior. | |||||||||||||||||||||||||||||||||
| 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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4.2 ★★★★★
Based on 933 reviews
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Product Reviews
★★★★★ 5
Still squeaking after all these days!
Size: Jumbo, Style: Beef
My power chewer GSPs usually wreck all "tough" toys in less than an hour, especially if they squeak. These seem pricey but I can say it's held up really well. After about three weeks, it's still hanging in there! The stripes are worn off but the squeaker still squeaks and that's what it's all about.
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Reviewed in the United States on May 15, 2026
★★★★★ 5
Long lasting scent and has not been destroyed by my heavy chewer.
Size: Jumbo, Style: Bacon
My medium puppy loves this ball. She already has the smaller size but appears to enjoy the challenge of catching the large one. An added benefit is it's easier for me to win it back when we fight (play fighting) for it.
The scent really lasts. I've had the smaller version for a few months and the scent is still there. Does it really smell like bacon? Not to my nose but the puppy thinks it's awesome.
The ball itself is really tough. The pup can destroy a tennis ball in a day. Except for a few teeth marks these balls are holding together great.
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Reviewed in the United States on June 10, 2026
★★★★★ 5
Worth every penny—my beagle is obsessed
Size: Medium, Style: Cheddar Cheese, Size: Medium, Style: Cheddar Cheese
No dog toy is indestructible if you have a hound or a pit. I happen to have a combination of both. This toy is the best $12 you could possibly spend for a dog that likes to “kill” their toys. The only reason I’m buying more is because I keep losing them.
My dog is a rescue. She’s half beagle, a quarter boxer, and a quarter pit bull. She’s a tiny fire-cracker and I lover her to pieces. And she loves tennis balls to pieces. Literally.
She adores tennis balls, squeaky toys, and food. She will absolutely try to surgically extract the squeaker from any toy she gets because she has a high prey-drive. These balls last usually at least a week before she manages to dislodge the squeaker or puncture the squeaking mechanism, but I still have one I got about six months ago that has the paint worn off and it still squeaks.
This ball literally has everything. It squeaks, it bounces pretty high, it looks like a real tennis ball so my girl recognizes them as “toy” immediately, and they smell good.
These balls in particular last forever. I got her a can of regular tennis balls and she had them in four pieces within 10 minutes or less, which is not healthy for her because she likes to eat the pieces. For this dog toy, I have never seen her crack it. She’s pushed the squeaker inside of exactly one of them so far, and it stayed inside the ball. Much safer than a toy that she can possibly ingest.
The toys come in several colors, but it’s not just to make them look cute and colorful. The colors correspond to flavors, as each of the balls has a different scent. I’ve sniffed them and noticed there’s a mild food smell beneath the rubber scent. I can tell that dogs really do get a kick out of the “flavors.”
I’ve taken these to the dog park and all the dogs want the Playology tennis ball, even if we have other generic balls flying around. My dog even has favorites. We have a muscle-head pit bull/staffy mix at the park who is a giant meat-ball. He got his hands on one of these and he wasn’t even able to break it open for at least 20 minutes of deliberate chewing.
My girl’s favorite “flavor” for a long time was beef (the red one). The blue “peanut butter” flavor is the easiest one to find in stores. She was obsessed with a purple one for a while, apparently it was bacon flavor. I couldn’t find that one! I saw that Amazon has an orange “cheese” flavor and knew she’d love it. She’s obsessed.
I really wanted to leave this review to convince people to invest in this product. Your dog will be so happy, and this company made something great.
I’m worried because I went on their website yesterday and they had only one or two colors available. I don’t want them to stop making this amazing toy. I hope they don’t lose business because their product is TOO durable!! If they make more flavors, or maybe make some multi-dog (tug ropes?) or similar, I’ll definitely continue to be a customer. Please keep making these awesome tennis balls!
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Reviewed in the United States on January 13, 2025
★★★★★ 5
Not Squeakable for Mini/Toy Aussies
Size: Medium (VALUE 2-Pack), Style: Chicken
Very durable and good squeaker. However, too large and hard for my Mini Aussies to squeak. My Aussies love their squeaky toys, unfortunately not this one.
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Reviewed in the United States on May 1, 2026
★★★★★ 4
Not indestructable, but very sturdy
Size: Medium, Style: Beef
I have recently been blessed (and cursed) with my first Super Chewer dog, a year old Australian Shepherd. She will chew herself out of boredom, so it's imperative we always have sturdy toys readily available for her, preferably in every room.
Playology and Kong by and large have been the best toys we can find for her. Kong has a very good rubber ball, bone, and ring that have held up spectacularly (no noticeable damage after months of use, except maybe some pin-prick teeth marks). Dogs need a wide range of textures though for enrichment, so we've been dabbling with the Playology line (the tire, plush bone, dry-tech rope), including this ball.
The scent factor is definitely interesting. Most people know Playology for the peanut butter scents, so I was very excited to find a multitude of flavors available on Amazon, especially with the different scents clearly labeled. I choose the red or beef flavored since the only two in the medium size in stock at the time were beef and sweet potato. The scent is indeed noticeable to humans, but it's not too strong or overwhelming on its own (walking down the Playology aisle in Petsmart can be overwhelming though since they have dozens of dozens of scented toys, much like going down a candle aisle).
I can't speak for the dog on how she feels about the scent, but she took to the ball immediately, same as the normal peanut butter ones we get. The medium size is pretty much perfect for her (she's just under 50lbs). It's got decent bounce for playing fetch in the house, but not so bouncy that we're afraid to use it in the house. It's got a decent density to it as well that adds to the sturdiness.
The biggest issue is the squeaker. It's not an obnoxious squeaker by any means, and the dog enjoys it very much. However, when you put a squeaker in any toy, it reduces the density and creates a weak spot dogs can exploit. She did good with the ball at first and was able to sit and chew on it for minutes as a time without doing any real damage (her teeth leave lots of pin-prick teeth marks, but they don't do any real damage to the ball, just superficial marks).
Aussies are considered incredibly smart though, and it didn't take her long to learn she could rip out the bottom of the ball where the squeaker is sealed inside to tear the squeaker out, thus ripping the squeaker apart and compromising the structural integrity of the ball. We're debating if we want to toss the ball in the trash or attempt letting her play with it in a supervised session without the squeaker to see if it is safe.
So, overall, this is a very tough ball. It does accumulate teeth marks that are essentially harmless, but there is a weak spot in the seam where the squeaker is inserted, which means this is not indestructible. It is by and large a great ball for fetch and likely won't ever be destroyed unless a dog is left with it long enough to chew a hole through that squeaker seam. Playology balls are pretty costly though, so the risk of it becoming unusable once the squeaker is ripped out is a bit off-putting. The bright side is that if you monitor your dog, you can likely get several weeks or months of use out of the ball; just no unsupervised play sessions.
7/10, durable, bouncy, and the scent keeps the dog interested, but the ball is not good unsupervised as it can be destroyed if the squeaker is ripped out. We will continue to buy Playology toys, but we will likely not buy too many of these balls until a more durable one (maybe without a squeaker) is released.
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Reviewed in the United States on March 20, 2022