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Description
Rat NHE3 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 a sodium–hydrogen antiporter 3 (NHE3) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by HRP peroxidase and to yellow by acid. The intensity of the color is positively correlated with the sodium–hydrogen antiporter 3 (NHE3) content in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Sodium–hydrogen antiporter 3 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Sodium/hydrogen exchanger 3 (NHE3), also known as sodium-hydrogen transporter 3 or solute carrier family 9 member 3 (SLC9A3), is a protein encoded by the SLC9A3 gene. It is a sodium-hydrogen antiporter. It is found in the apical region of proximal tubular epithelial cells of the kidney, in the apical region of enterocytes, and in the basolateral region of duodenal and pancreatic cells, responsible for releasing HCO₃ into the duodenal lumen. It is primarily responsible for maintaining sodium homeostasis and is also indirectly involved in buffering blood pH. | |||||||||||||||||||||||||||||||||
| 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, cell lysates, and other biological fluids |
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4.1 ★★★★★
Based on 281 reviews
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Product Reviews
★★★★★ 5
Hotel quality, budget friendly
Size: King, Style: Light
I’ve only used this for several nights. Overall I’m pretty happy with it, especially for the price. It is soft to the touch and has a nice, smooth feel. It’s lightweight but still provides enough warmth for mild to cool nights. I don’t like feeling weighed down when I sleep, so the light fill is actually a big plus for me. If you’re someone who sleeps hot, this is a solid choice. The corner tabs are super helpful. I paired it with a duvet cover and haven’t had issues with shifting or bunching. It also fits my bed well, though I could see it being a bit short on the sides if you have a very thick mattress. This is a great budget-friendly option and seems like it will be easy to maintain.
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Reviewed in the United States on September 19, 2025
★★★★★ 5
A Great Comforter for Everyday Use
Size: Full/Queen, Style: All-Season
I absolutely love this comforter! I love the fact it’s machine washable, and it held up really well after the first wash. It fluffed back up nicely, stayed soft, and didn’t lose its shape. It’s not too heavy but still cozy enough to keep you comfortable. For the price, the quality exceeded my expectations, and I am very happy with this purchase. I highly recommend 😊
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Reviewed in the United States on June 4, 2026
★★★★★ 5
Excellent down alternative insert
Size: Full/Queen, Style: All-Season
I purchased this down alternative for my Airbnb because we can easily wash it. I love that it is fluffy and warm and easy to put into the duvet. Price was great as well!!
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Reviewed in the United States on April 19, 2026
★★★★★ 5
Feels luxury, I approve
Size: King, Style: Warm
I have purchased high end comforters/duvet inserts and this one is somehow my favorite. It's so thick and fluffy.
I was going to do the double duvet insert method to create an extra lush bed situation, but honestly it's not needed at all with this comforter! It poofs up wonderfully inside my duvet cover (king duvet, king insert) and it's like heaven. It's very cozy yet airy, and it moves well during the night and does not fee too heavy despite its weight.
This thing is definitely on the warm end and probably won't be suitable for Florida summers, but depending on how you set the AC at night it could be fine!
The comforter is weighty, it feels like quality, and has a soft fabric. I wouldn't use it without a duvet cover, that's more of an aesthetic choice but I like keeping it clean. The white is very white.
Honestly, I will be getting another one simply because I'm so impressed. Definitely no need to drop $300+ dollars elsewhere.
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Reviewed in the United States on October 25, 2025
★★★★★ 4
ALL SEASON IS TOO WARM!
Size: King, Style: All-Season
I got the King, All Season, to squeeze into a Queen duvet for extra floof and it's sooo cozy. Highly recommend.
I redacted 1 star though because although it says All Season, the amount of down in here feels like it's for winter. The first night I used it I woke up dripping in sweat. It's currently March, I live in Los Angeles area, hitting about 75° weather outside rn, but I sleep with AC full blast at night (usually hit about 64 when I wake up). I can't sleep with it, I'll need to buy the "cool" one, and hope that isn't tooo thin.
Don't get me wrong, it's a great insert, but holy hell, it's too heckin' warm. I saw someone else mention it and wish I had honed in on their comment. I don't wanna return it as it was used, so I'm stuck with it now.
*UPDATE*
I am back because I also bought the "Cool" blanket as well after experiencing the heat of the all season and it too is also too hot for me. I've had a duvet before but I guess not filled to the capacity of these blankets. If you're a hot sleeper maybe look elsewhere?
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Reviewed in the United States on March 12, 2026