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Description
Rat 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 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 a capture antibody against ATPase, Na+/K+ transporting, alpha 1 (ATP1a1). After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by peroxidase (HRP) catalysis and to yellow by acid. The intensity of the color is positively correlated with the presence of ATPase, Na+/K+ transporting, alpha 1 (ATP1a1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat 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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4.2 ★★★★★
Based on 6 reviews
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Product Reviews
★★★★★ 5
Mind blowing page turner
Oh my stars!!! 🤯 he falls first and he falls hard, enemies to lovers, fated mates. Every characters are lovable. There's action, magic, one horse 🤭 and a plot twist that keep on twisting. Oh and a bad ass FMC! Loved it!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 23, 2026
★★★★★ 3
Interesting. Confusing ending that's too similar to another book
Ok, I had to process almost a whole 24 hours before I could write this review. And long story short, did I enjoy the book, yes. Does it have a lot of interesting and good parts to it? Also yes. But does it also have some major flaws? Absolutely.
Now I'm not gonna break down every single little thing in this book. But here are the basics of what I liked, and what I didn't like.
The good? I liked the characters. They intrigued me off the bad. The world building is pretty decent. It's a little confusing in the beginning, but information is slowly doled out, and some questions are answered. I found that there was a lot of little twists and turns that kept the story engaging. The magic system is intriguing.
But, there are quite a few things off for me. First off, we have another story that has heavy inspirations from other books. There are a lot of aspects in this story that felt directly pulled from throne of glass. And the big twist at the end? Was almost verbatim the same ending as the book how does it feel. As soon as I read it I was like hold up, I literally just read almost this same thing when I read how does it feel when it released like a year or year and a half ago.
Now I know no concepts are really new anymore, and inspiration comes for everywhere. But I feel like most stories it's like, oh this book has these vibes, or if you liked this book you'd like this one that's similar. But this reminds me of powerless in where there are like exact plots and plot points taken from other things. Now is that a bad thing? Not necessarily. I still enjoyed powerless, and I still enjoyed this story. But it does throw me.
There are also a few red flags that the MMC Acker gave me that were not the good kind of red flags we love. First, when they are riding thru the city and people throw stuff at the FMC and he does nothing? Red flag. When they meet his dad and they demand she vows not only to the king but to the MMC? Red flag. All the secrets he keeps? Red flag. When he SLAPS HER IN THE END? Red flag. I'm not sure how I really feel about him.
In the end. This was still an enjoyable read. I did like it and I am curious about the next book. But I am wary about some of the plot points and the MMC.
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Reviewed in the United States on November 26, 2024
★★★★★ 5
Just read it. You won’t regret it.
Format: Hardcover
This book completely blew me away. I loved every single character—not just the FMC and MMC, but the side characters too, which is rare. In a first book, side characters can feel flat while the focus is on world-building, but that definitely wasn’t the case here.
The romantic tension between the FMC and MMC was so good—seriously addictive. The plot itself felt pretty straightforward at first, and the world-building was easy to follow. It even gave me Heartless Hunter / Wolf King vibes… until the last 20 pages.
And wow—those final pages. I usually spot plot twists coming, and I thought this one was going to be predictable, but I was completely wrong. That ending genuinely shocked me—I haven’t had that reaction to a book in a long time.
I’m already counting down to book two (and that teaser chapter? Absolutely brutal).
Just read it. You won’t regret it.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 4, 2026
★★★★★ 4
4 Stars for Now!
Format: Kindle
I DEVOURED this book. I was in a huge slump after reading a horrible series right before this but after 3 pages of this book, I was hooked. I read the whole book in 2 days. I think the author did an amazing job at writing an engaging story with an amazing twist at the end. I know many readers feel betrayed by the twist and find the narrator to be unreliable but I feel that the twist was done in a great unsuspecting way. Though I do feel betrayed, I think there is so much room for redemption in the second book!
The author also did an amazing job at world/character building. She was able to do so without interrupting the story which is one of my huge pet peeves. A lot of authors build by taking breaks in the plot or dialogue which I find choppy and disengaging but this author was able to build in an engaging way. There was some lack of world/character building but I think that was due to keeping reader in the dark. The world, politics, and characters were all very easy to understand.
I found the romance a little lackluster and a bit off-kilter. Some of it was really pushy and rushed which left a bad taste in my mouth. I do hope the author delves deeper into the romance in coming books.
The FMC was a breath of fresh air. She was not whiny or stuck-up like most books. She was never afraid to accept help from others but she was still able to shine with her intelligence. Not once did I think she made a poor decision just for the sake of a plot point. She was really easy to like. The MMC could use some work but I think that will happen more in coming books.
This is a MUST read, my only advice would be to wait until it is a finished series!
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Reviewed in the United States on July 30, 2025
★★★★★ 5
Need book 2 please :-)
Format: Kindle
From the very first chapter, Metal Slinger hooked me and refused to let go.
Set in a brutal, metal-bound world where power is carved from steel and survival depends on strength, strategy, and sheer will, this story follows a young woman forced into a dangerous system where control over metal isn’t just magic — it’s dominance, protection, and rebellion. When she’s pulled into escalating conflict and shifting alliances, she quickly learns that power always comes with a cost.
The magic system is sharp and immersive without being overwhelming. The way metal manipulation is woven into combat and politics makes every confrontation feel cinematic and high-stakes. It’s gritty, tense, and beautifully layered.
The Brynn, our FMC absolutely shines. She’s fierce but not untouchable, resilient without feeling unrealistic. Watching her navigate betrayal, survival, and the slow awakening of her own strength felt earned every step of the way. There’s grit in her fight scenes and vulnerability in her quieter moments that make her deeply human.
And the tension? Immaculate. The chemistry simmers under the surface before sparking into something electric. Every reluctant alliance, every charged interaction, every moment of “are they about to destroy each other or kiss?” had me flying through pages.
The pacing is relentless — no filler, no wasted scenes. Twists land hard. Stakes keep climbing. And that ending? Absolutely feral. The kind that leaves you staring at the wall for a minute before immediately checking when book two releases.
✨ Gritty metal-based magic
⚔️ High-stakes survival
🔥 Slow-burn tension
🖤 Fierce, layered heroine
💥 A finale that HITS (but cliffhanger!)
This is one of those fantasies that reminds you why you fell in love with the genre in the first place. If this is just Book One, I am more than ready to be emotionally wrecked again in the sequel.
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Reviewed in the United States on February 14, 2026
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