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Description
Human ATP1a1 ELISA KitProduct Specification Usage Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High precision pipette and gun tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37 constant temperature box 4. Distilled water or deionized water Sample processing and requirements: 1. Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4 overnight, then centrifuge at 1000g for 20
Product Specification
| Usage |
Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High-precision pipette and gun tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37℃ constant temperature box 4. Distilled water or deionized water Sample processing and requirements: 1. Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4℃ overnight, then centrifuge at 1000×g for 20 minutes, and take the supernatant, or store the supernatant at -20℃ or -80℃, but avoid repeated freezing and thawing. 2. Plasma: Collect the specimen using EDTA or heparin as an anticoagulant. Centrifuge the specimen at 1000 × g for 15 minutes at 2-8°C within 30 minutes of collection. The supernatant can be assayed or stored at -20°C or -80°C, but avoid repeated freezing and thawing. 3. 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 the tissue and mince it. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1 g of tissue sample to 9 mL 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 the supernatant can be assayed. 4. Cell Lysis Buffer: Gently wash adherent cells with ice-cold PBS, then trypsinize and collect cells by centrifugation at 1000×g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash collected cells three times with ice-cold PBS and resuspend 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 freeze-thaw cycles or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and remove the supernatant for analysis. 5. Cell Culture Supernatant: Centrifuge at 1000×g for 20 minutes. Remove the supernatant for analysis or store at -20°C or -80°C, avoiding repeated freeze-thaw cycles. 6. 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 competitive enzyme-linked immunosorbent assay (ELISA). Samples, standards, biotin-labeled antibodies, and HRP enzyme conjugates are added sequentially to microwells pre-coated with the universal species ATPase, Na+/K+ transporting, alpha 1 (ATP1a1) antigen. After incubation and washing, the sample is developed with the substrate TMB. TMB is converted to blue under the catalysis of peroxidase (HRP) and to the final yellow under the action of acid. The depth of the color is negatively correlated with the universal species ATPase, Na+/K+ transporting, alpha 1 (ATP1a1) in the sample. The absorbance (OD value) is measured at a wavelength of 450nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human ATPase, Na+/K+ transporting, alpha 1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Sodium/potassium-transporting ATPase α1 peptide, also known as ATP1a1, is an enzyme encoded by the ATP1a1 gene. The protein encoded by this gene belongs to the P-type cation-transporting ATPase family and the Na⁺/K⁺ ATPase subfamily. Na⁺/K⁺ ATPase is an integral membrane protein responsible for establishing and maintaining electrochemical gradients of sodium and potassium ions across the plasma membrane. These gradients are crucial for osmotic regulation, sodium-coupled transport of various organic and inorganic molecules, and electrical excitability in nerves and muscles. The enzyme consists of two subunits: a large catalytic subunit (α) and a smaller glycoprotein subunit (β). The catalytic subunits of Na+/K+-ATPase are encoded by multiple genes. This gene encodes the α1 subunit. Alternatively spliced transcript variants encoding different isoforms have been identified. In melanocytes, ATP1A1 gene expression may be regulated by MITF. Mutations in this gene are associated with aldosterone-producing adenomas and secondary hypertension. | |||||||||||||||||||||||||||||||||
| 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 | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.0 ★★★★★
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Product Reviews
★★★★★ 5
beautiful sapphic second chance
Format: Kindle
I could not put this book down, I devoured it in one day! A heartfelt sapphic romance that pulled you in from the start. Gemma and Caitlin’s dual POVs and dual timelines was done effortlessly. Seeing the struggles of a high school romance was so relatable. Then flash forward to the present when they were forced into each other’s lives after nine years because they were cast in the same movie! The cross between past, present, and the movie being filmed was full of angst, longing, regret, and hit me on every emotional level.
I could not recommend this book more!
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Reviewed in the United States on January 22, 2026
★★★★★ 3
Missed the Mark *Spoilers*
Format: Kindle
I was surprised by the author’s bio because at times it felt like this book was written by an adolescent imagining what filming a movie would be like. I really enjoyed certain parts of the book, especially the flashbacks to HS, the flirtatious note-writing and the botched Spanish terms of endearment. But the main feeling I had reading it was whiplash, in terms of all the back-and-forth between the MCs. Even at almost 30 years old, they both felt emotionally immature.
**Spoilers Ahead**
It was odd to me that when they finally confronted their past, it was in terms of the summer after their sophomore year of HS, rather than the big breakup when they were in college. Skipping ahead, the Paris ending felt very abrupt and out of character.
I also felt that certain side characters (Troy, Darbie, Emily, etc.) were very two-dimensional and discarded easily where it would’ve been interesting to see their reactions to the MCs’ relationship.
I can see where the author laid the groundwork for a good series, it definitely has potential. This book just missed the mark for me.
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Reviewed in the United States on February 17, 2026
★★★★★ 4
Fantastic Second Chance Romance
Format: Kindle
The only reason this book wasn’t a 5* was because Amazon doesn’t allow 4.5* and I felt like parts of the rekindling of their romance at the very end were slightly rushed. Aside from that, this book was INCREDIBLE. I fell in love with them as a couple from the beginning. Gemma’s back and forth emotional parkour was a little rough at spots with how quickly she changed from wanting nothing to do with Caitlin to wanting to take her home. BUT I loved the epilogue and the full circle moments of them together and their future. Can’t wait to read another book by this author!
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Reviewed in the United States on February 7, 2026
★★★★★ 5
Must buy
Format: Paperback
Support any lgbtq writer
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Reviewed in the United States on March 26, 2026
★★★★★ 5
these are her viewpoint on a large mix of poems ... The Final Work
Format: Hardcover
helen vendler has done a magisteral job of presenting her poem selections
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Reviewed in the United States on April 29, 2026