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Description
Human ATP1b4 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 ATPase, Na+/K+ Transporting Beta 4 Polypeptide (ATP1b4) 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 ATPase, Na+/K+ Transporting Beta 4 Polypeptide (ATP1b4) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human ATPase, Na+/K+ Transporting Beta 4 Polypeptide ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | The protein ATP1B4 (ATP1b4), also known as ATPase Na+/K+ transporting family member β4, is encoded by the ATP1b4 gene. It functions as a transcriptional regulator during muscle development through interaction with SNW1. It has lost its ancestral function as the β subunit of the Na,K-ATPase. Diseases associated with it include mannoses, Beta A, lysosomal, and thyrotoxic periodic paralysis. Pathways involved include GPCR signaling and the transcriptional activity of the SMAD2/SMAD3-SMAD4 heterotrimer. Two transcript variants encoding distinct isoforms have been identified for this gene. | |||||||||||||||||||||||||||||||||
| 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 | 0.312-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates and other biological fluids |
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4.6 ★★★★★
Based on 25 reviews
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Product Reviews
★★★★★ 5
Excellent vacuum!!
Color: White
This is a hard working little robot. It cleans dirt and debris so easily and even gets the Arizona dust that accumulates everyday. I have a cat the sheds and 4 boys plus my husband and this vacuum works like a champ. I haven’t tried the mop feature yet because I just haven’t sat down and read the directions. I’ll update when I try that feature out. I love that it can go under my furniture and climb up rugs, even my cat scratch boards. I 100% recommend this vacuum. It has changed my house keeping routine and saves me a ton of time. I absolutely love it and you can’t beat the price!
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Reviewed in the United States on November 15, 2025
★★★★★ 5
Pleasantly surprised.
Color: White, Color: White
Based on the price I wasn't expecting much but boy was I wrong. I'm so impressed and thrilled with this purchase. I have a long haired Maine Coon and it picked up so much fur. It picked up thread, small pieces of yarn and feathers from my pillows. I watched the long front brushes pull dust and dirt out from under spaces that my regular vacuum would never get without moving things. I have carpet, hardwood, throw rugs and even a long shag throw by my bed that it maneuvered over mostly easily. I helped it over one or two transitions but it never got stuck. Just turned and kept going. The mop worked great. Only concern is that it only uses water so it's not a deep clean but the pad was dirty and once it dried I threw it into the wash. Definitely will use it in between a deeper cleans making cleaning a whole lot easier. With a cat shredding the carpet on a scratching post and all of his fur I was shocked how packed the dirt cup was and how great the house looked with all the little bits of scratching pole, oh yeah and card board scratching pad cleaned up so easily. I highly recommend. The cat thinks it's a cat toy so he kept starting it up. I had to cover it to protect it from the cat. It now lives under it's very own plastic bin. 🙂
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Reviewed in the United States on March 13, 2025
★★★★★ 5
Robotic Vacuum
Color: Jet Black, Color: Jet Black
Good Cleaner
I'm very old and it was very difficult for me to clean my home but thank God my Daughter told me about Robotic Vacuum cleaners so, I ordered it and now I'm using it, really impressive results.
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Reviewed in the United States on April 22, 2026
★★★★★ 5
My first experience with a Robot vacuum
Color: White
I am liking this robot vacuum. It is very easy to program and can be done from your phone. It has mapping capabilities which helps with learning the room. The remote is easy to use too. It picks up most dust particles and pet fur from rugs and bare floors. It has a wet floor mopping pad which is so helpful. Surprisingly it picks up more than you think. The dust cup holds quite a bit before you have to empty it. The little side brushes help with collecting the particles. It can go under some furniture and along the walls. You have several floor patterns that it can be programmed for. It is very quiet too. Recharging doesn’t take long either. It can even be programmed to vacuum while you are away. Instructions are downloaded through the app.
My cats are very curious about how it moves about the rooms. So much fun to watch them.
I really got a great bargain for this robot vacuum on Amazon.
(I found that if you have clutter on the floors or cords under furniture you should remove and secure them before vacuuming.) It will find a way to go from floor to area rugs once it maps out the area.
I am still learning the functions in the app👍🏾👍🏾😃
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Reviewed in the United States on January 9, 2025
★★★★★ 1
Piece of junk
Color: White
So many problems with this device. I have a one bedroom apartment with one bathroom and no carpet so it's all hardwood floor. You would think this device would be able to vacuum the entire place without problems right? I think it intentionally looks for places to get stuck. For instance I have electrical plugs underneath my desk pushed against the wall, and somehow it managed to get stuck in them. So I had to block that area with a box. Then it got stuck going over my bath mats. So when I vacuum I have to put them into the shower. Then somehow it managed to get stuck in a little area by the island in my kitchen. So I had to put a roll of paper towels there so it wouldn't go into that area. Then it got stuck under the wheels of my desk chair. So I had to block that area. One night, even though there's a big table on the side of my couch to block it, it figured out a way to get behind my couch and then stopped. It took me two days to find it. And that's the other thing. it doesn't have enough power to clean an entire one bedroom apartment. So even if it doesn't get stuck, I will find it in the middle of the floor with the battery drained.
When I watched it I noticed that it will bang something and then it'll keep banging it a few times before moving on. And by moving on I mean it'll move 6 in and bang there a few times before moving on again. That's probably how it got stuck under the island. It couldn't fit the first time, so it kept pushing until finally it got jammed in there. I must say, however, it is amusing when it bangs into my foot and doesn't give up and tries to climb over my foot.
So although I bought this thing the end of November, I was in the hospital and wasn't able to start using it until the third week in January. After a couple of weeks I wanted to return it, but unfortunately I can't, because the window to return it ended January 31st.
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Reviewed in the United States on February 16, 2025