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Description
Human ADRA1A 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 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, biotin-labeled detection antibody, and HRP conjugate are sequentially added to microwells pre-coated with an Adrenergic Receptor Alpha 1A (ADRA1A) 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 Adrenergic Receptor Alpha 1A (ADRA1A) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Adrenergic Receptor Alpha 1A ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | The gene encoding the adrenergic α1A receptor (ADRA1A) is called ADRA1A. A subtype of the receptor, α1C, has been discovered, which is identical to the previously discovered α1A receptor subtype. ADRA1A binds to both epinephrine (adrenaline) and norepinephrine (noradrenaline). Once activated, it promotes cell growth and proliferation. | |||||||||||||||||||||||||||||||||
| 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, cell lysates, and other biological fluids |
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4.6 ★★★★★
Based on 22 reviews
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Product Reviews
★★★★★ 5
Great for my pitbull, worth the price! the warranty isn’t what it seems so check their site first.
Color: Orange Squeak Toy, Size: Medium
I’ve now purchased 3 and I don’t see myself ever stopping! My 55lb Staffie (pitbull) has absolutely obliterated every ball and toy (rope, rubber, stuffed) before this one in a matter of minutes. You take your eyes off it, and the next time you’d look back it would be in pieces. Kong Xtremes and other “indestructible” toys only lasted a week, tops. He also goes nuts for anything that squeaks, so keeping him happily entertained for a while was a challenge because the second he’d chomp down on a squeaker, it’d break. He’s now had this blue squeak ball for a MONTH and it STILL SQUEAKS!! He regularly falls asleep with it still in his mouth, or tucked under his chin. That’s how obsessed he is with this thing. I would 100% recommend this to anyone who needs a seriously durable toy for their super powerful chewer as that’s been my experience! This material is super bouncy and he can bite down (unlike some rubbers, Kong/West Paw) without puncturing it somehow. It’s perfect for fetch, and floats too. He’s never had a toy last this long, so this is a welcome break for my wallet despite being a more expensive toy. I was buying a few toys every week until we tried this one!
Cons: The first two balls didn’t squeak after a few days, so maybe they’ve improved something since then. The warranty replacement process is more tedious than others like West Paw, and only replaces them ONCE. It’s also requires more force to squeak this given the nature of the material, but since it extends the life of the squeaker I’m totally fine with it.
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Reviewed in the United States on December 13, 2018
★★★★★ 5
This ball has lasted and still have it after many months or longer. My dog loves it.
Color: Pink Squeak Toy, Size: Medium
My Belgian Malinois can really chew through inferior play toys and balls, etc., and this is one of the few that has lasted a long time, and he really likes it. Another bonus is that it gets quite slippery with his saliva and the ball will bounce a little bit unevenly, so it can either slip out of his mouth or bounce in an unexpected direction that can fool him. He has such quick reactions that is hard to get a ball past him, but with this ball, I can score extra points on him when he can't block it. Otherwise I have no chance, and he beats me every time.
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Reviewed in the United States on October 21, 2025
★★★★★ 3
Please go back to the original ball design and squeaker!!
Color: Pink Squeak Toy, Size: Medium, Color: Pink Squeak Toy, Size: Medium
Review update: I am updating my review for a few reasons. 1) The ball I ordered below was the 3rd time I purchased it. I originally bought one years ago, Pre-Covid, for my old pitty. My now Corgi, Jett loved that ball so I got a 2nd one for times when we couldn’t find the original. Well, the ball was slightly larger, the squeaker stopped working, and Jett preferred the original. 2) The newest one doesn’t squeak at all and is also slightly larger than the original. The original fell down the sewer drain that is in our backyard, and we lost the 2nd ball, so we ordered the ball again. My husband ended up having to retrieve the original from the sewer drain because Jett would pace around the sewer looking for the original. I ended up returning the 3rd ball because Jett doesn’t want anything to do with it & it doesn’t squeak. The video I provided is Jett squeaking the original that is at least 5-6 years old. I highly recommend the company go back to the original design that has not only stood the test of time and heavy use but is also the preferred size and for some reason has a much better, more reliable squeaker.
I bought 2 of these balls for my Corgi mix and another for my son’s girlfriend’s Golden Retriever. Both dogs love the ball. They love the squeak and the ball is very durable. The ball doesn’t easily squeak by hand but will when they bite down on the ball or when it bounces. Speaking of, it has a decent amount of bounce to it too. Our dog loves to jump in the air to catch it after it bounces off the ground. I highly recommend this ball for medium and large dogs.
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Reviewed in the United States on December 24, 2023
★★★★★ 5
Chuckit! Max Glow Ball Dog Toy - A Canine Delight in the Dark!
Size: Small, Pattern Name: Pack of 1
I recently introduced the Chuckit! Max Glow Ball Dog Toy to my furry friend, and it has quickly become a favorite in our playtime routine. Here's why it deserves a glowing 5-star review:
**Pros:**
**1. Day and Night Fun:**
The Max Glow feature of this ball adds an extra layer of excitement to playtime. Whether it's a sunny day or a moonlit night, my dog can enjoy fetching and playing with this ball anytime, making it a versatile and engaging toy.
**2. Long-Lasting Glow:**
The glow effect lasts surprisingly long, providing extended play sessions even after the sun sets. The consistent glow adds a thrilling element to our nighttime activities, making it a standout feature of this toy.
**3. Durable Design:**
The Chuckit! Max Glow Ball is built to last. Its durable construction can withstand the enthusiastic chewing and fetching antics of my dog, ensuring longevity and making it a reliable addition to our collection of toys.
**4. Easy to Clean:**
Cleaning this ball is a breeze. Whether it's muddy paw prints or slobber, a quick rinse under the tap is all it takes to keep the Chuckit! Max Glow Ball looking and feeling fresh for the next round of play.
**5. Compatible with Chuckit! Launchers:**
The ball is compatible with Chuckit! launchers, adding an extra layer of fun to our play sessions. The launcher allows for longer throws, providing a good workout for my dog and adding an interactive aspect to our outdoor activities.
**6. Fetching Favorite:**
My dog absolutely loves fetching this ball. The texture and size make it easy for him to carry in his mouth, and the glowing feature captures his attention and keeps him engaged during playtime.
**7. Versatile Outdoor Toy:**
Whether it's a game of fetch in the backyard, a trip to the park, or an evening stroll, the Chuckit! Max Glow Ball has become our go-to outdoor toy. Its versatility and engaging qualities make it an essential part of our canine adventures.
In conclusion, the Chuckit! Max Glow Ball Dog Toy has brought endless joy and excitement to our playtime routine. Its durable design, long-lasting glow, and compatibility with Chuckit! launchers make it a 5-star favorite in my dog's toy collection. If you're looking to add a touch of magic to your dog's play sessions, this glowing ball is a must-have. Let the nighttime playtime adventures begin! 🌙🐾🌟
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Reviewed in the United States on January 26, 2024
★★★★★ 5
I've bought like 20! Love them!!!
Size: Large, Pattern Name: Pack of 1
My dog is obsessed with this ball! Glow in the Dark is super bouncy, easy to spot at night, and tough enough to handle rough play. Perfect for evening fetch sessions—fun for both of us! We have bought about 20 of these so far. I always bring my dog to play fetch at night, and this is perfect if you're playing in a field or beach. Glows in the dark so both you and your dog can see it. The size of the ball is great for my big dog. It is also the perfect type of silicone so that my dog cannot easily tear it apart and chew it, which is GREAT! We do NOT want our dog's ingesting silicone at all. If you have a ball chewer, this ball is perfect and durable. Chewing ability is good and is not too tough but tough enough.
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Reviewed in the United States on August 10, 2025