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Description
Human NKCC2 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 Na-K-Cl Cotransporter 2 (NKCC2) 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 amount of Na-K-Cl Cotransporter 2 (NKCC2) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Na-K-Cl Cotransporter 2 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Sodium-potassium-chloride cotransporter 2 (NKCC2), also known as solute carrier family 12, member 1 (SLC12A1), is a protein encoded by the SLC12A1 gene. It is specifically found in the thick ascending limb of the loop of Henle and in the macular densa cells of the nephron, the basic functional unit of the kidney. Within these cells, it resides in the apical membrane adjacent to the nephron lumen, the hollow space containing urine. Thus, it functions both in sodium absorption and in tubulospinal feedback. | |||||||||||||||||||||||||||||||||
| 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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4.3 ★★★★★
Based on 14 reviews
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Product Reviews
★★★★★ 5
Absorbent and Make Cleaning Easier
I purchased this Scott paper towel pack for everyday household cleaning, and overall, they have worked very well for daily use. They are practical, easy to store, and convenient for families that go through paper towels frequently.
One of the biggest advantages is the pack size. The family-size bundle lasts a long time, and I like that the rolls are packaged into two separate bundles inside the larger pack, which makes storage and organization easier. The rolls themselves are slightly smaller in width, which actually works well because they fit easily onto any paper towel holder without being bulky or difficult to spin.
In terms of performance, these towels are surprisingly absorbent despite being thinner than some leading brands. The ridged texture on each sheet helps pick up dirt, spills, and grime more effectively, and the “select-a-size” feature is very convenient since you can use smaller sheets when you only need a little. One of my favorite features is that they are lint free, which makes them especially useful for cleaning glass, mirrors, counters, and other surfaces where leftover fibers can be annoying.
There are a couple of minor downsides. The paper towels are definitely thinner compared to other leading brands, and the overall roll size is somewhat smaller as well. However, they still absorb a good amount of liquid and hold up well enough for normal household cleaning tasks.
Overall, these are a reliable and convenient everyday paper towel option. They may not be the thickest on the market, but the absorbency, lint-free design, and practical roll size make them a great choice for regular use around the house.
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Reviewed in the United States on May 29, 2026
★★★★★ 5
Good paper towels.
At the time of my review this set of 16 double layered paper towels are going for about $18.99. Not a bad price for a namebrand paper towel.
I usually use Great Value paper towels because they are not only inexpensive but have good absorbency so it was interesting to compare it to the Scott namebrand, whom I use just about exclusively for my toilet tissue.
The papertowels worked well as I expected them to but I did not specifically see a dramatic difference between it and the ones I usually used. The only noticeable differences to me were:
- Scott paper towels had a prettier pattern
- Scott paper towels visually had a much more tidy design and *looked* higher quality
- Scott paper towels felt softer to the touch
As for absorbency though? I felt like they performed identically. And trust me, we burn through paper towels. We have two small children and still have to wash bottles daily. So, we try to balance cost versus efficacy for papertowels because we use them so much every day.
For the what they are these are great, but I will probably stick to the other brand unless I'm in a pinch and need to order papertowels more conveniently OR if these ever go on sale or have a coupon. I don't need my papertowels to look or feel pretty (the sloppy floral print and more coarse feel of the GV ones work for us right now because they are used exclusively for clean up), I need them to absorb and pick up gunk well.
Would I still recommend these, though? Absolutely. Maybe appearance and texture are more important to you for a paper towels. We aren't there yet with our littles. Lol.
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Reviewed in the United States on May 11, 2026
★★★★★ 1
Junk
Just pay for the better ones, these are cheap junk. They can’t even be torn off without falling apart.
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Reviewed in the United States on May 30, 2026
★★★★★ 5
MY DOG LOVES IT
Size: 20.0"L x 20.0"W x 6.0''TH, Style: Round, Pattern Name: Bed
MY CHIHUAHUA WON'T GET OUT OF THIS BED! she loves it! from the first moment we brought it in and put it on the floor. she gently scratches at the sides and cuddles in with her blankey and likes she's not there. ohhh... great value for the price! nice size, big enough for both of my chihuahuas to lay in comfortably. soft, but very sturdy sides, the bottom is fluffy, padded very well. the outside bottom, there is a non stick covering so the bed doesn't scootch around on tile floor, it stays put.
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Reviewed in the United States on January 28, 2026
★★★★★ 5
It's lasted
Size: 20.0"L x 20.0"W x 6.0''TH, Style: Round, Pattern Name: Bed
I've had this bed for 5 years and it is still in great shape. Washes up in the washing machine just fine. My littlest dog absolutely loves it and is lost when our other bigger dog steals it. She barely fits in it but she loves it too. Ha! Good quality at a good price. Couldn't ask for more.
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Reviewed in the United States on May 27, 2026