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Description
Mouse 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 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 sodium/hydrogen exchanger 3 (NHE3) capture antibody. 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 sodium/hydrogen exchanger 3 (NHE3) content in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Sodium/Hydrogen Exchanger 3 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Na+/H+ exchanger 3 (NHE3), also known as sodium-hydrogen transporter 3 or solute carrier family 9 member 3 (SLC9A3), is a protein encoded by the NHE3 gene. It is a sodium-hydrogen antiporter. It is found in the apical region of proximal tubular epithelial cells of the renal tubules, in the apical region of enterocytes, and in the basolateral region of duodenal and pancreatic cells, responsible for releasing HCO−3 into the duodenal lumen. It is primarily responsible for maintaining sodium homeostasis and is also indirectly involved in buffering blood pH. It imports a sodium ion into the cytoplasmic matrix of renal tubular cells while simultaneously ejecting a hydrogen ion from the cell into the lumen of the proximal tubule. The sodium within the tubular cells may then be retained by the body rather than excreted in the urine. It indirectly contributes to blood buffering capacity because the ejected hydrogen ion is a product of carbonic anhydrase, which also produces bicarbonate. It has been shown to interact with CHP. | |||||||||||||||||||||||||||||||||
| 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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4.8 ★★★★★
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Product Reviews
★★★★★ 5
Great little speakers
Size: 5.25” Woofer, Size: 5.25” Woofer
These make great surround speakers, and they just sound good
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 22, 2026
★★★★★ 5
Good Speakers - Good Reputation - Good Purchase
Size: 5.25” Woofer
Good sounding, high quality speakers. These are Klipsch...enough said.
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Reviewed in the United States on March 31, 2026
★★★★★ 5
Klipsch R50 M next generation
Size: 5.25” Woofer
In spite of small size sound is very nice iam using it with a subwoofer and with my yamaha A4A AV receiver i like it iam hearing mostly heavy metal and the speakers have a nice punch and attack
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Reviewed in the United States on February 18, 2026
★★★★★ 4
compact surrounds that blend well with the right setup
Size: 4” Woofer
I picked up the Klipsch R‑40M as part of a full 5.1 system built around the R‑50M fronts and the R‑50C center. My original plan was to use another pair of R‑50M speakers in the back, but they simply didn’t fit in the rear cabinet space I have. The R‑40M ended up being the only model that fit comfortably, so they became my surround channels.
In terms of build quality, they feel solid and consistent with the rest of the Reference Next‑Gen line. The copper woofers and updated horn design give them the same visual identity as the larger models, which helps the whole system look cohesive. They’re compact enough to place in tighter spaces without compromising too much on performance.
Sonically, the R‑40M does a respectable job as a surround speaker. Effects are clear, directional cues are easy to follow, and they integrate smoothly with the more powerful R‑50M fronts. That said, they are noticeably less powerful than the R‑50M, so you need to spend a little time balancing your levels. Once I bumped their channel trim a bit and adjusted the distance settings, the rear soundstage locked in nicely and felt much more even.
They won’t deliver the same fullness or dynamic punch as the larger models, but for rear channels that’s usually not a deal breaker. What matters is clarity and consistency, and the R‑40M handles both well. Movies with active surround mixes feel immersive, and ambient effects wrap around the room without calling attention to the speakers themselves.
If you’re building a system where space is limited in the back, the R‑40M is a practical and capable choice. Just be prepared to fine tune your calibration so they match the output of the bigger speakers up front. Once dialed in, they round out a 5.1 setup very effectively.
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Reviewed in the United States on January 22, 2026
★★★★★ 5
Great Bang for the Buck!
Size: 5.25” Woofer
These R50 Studio Monitors are worth every penny. One could easily spend a heck of a lot more to get less. Have been a pro audio engineer my entire life. These monitors kick out clean, solid bass and accurate highs. Klipsch did a excellent job designing these monitors. Perfect for smail spaces where fidelity is critical. If you wall mount these, make sure you leave at least a 1.5 inch air gap between the back of the monitor and the wall. These units are sweet.
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Reviewed in the United States on July 22, 2025
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