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Description
Human NKCC1 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 1 (NKCC1) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue under the catalysis of HRP and to yellow under the action of acid. The intensity of the color is positively correlated with the amount of Na-K-Cl Cotransporter 1 (NKCC1) 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 1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Sodium-potassium-chloride cotransporter 1 (NKCC1) is a membrane transport protein. It is widely distributed throughout the body, particularly in fluid-secreting organs known as exocrine glands. Within the cells of these organs, it is typically located in the basement membrane, the portion of the cell membrane closest to blood vessels. Its basolateral location enables it to transport sodium, potassium, and chloride from the blood into the cell. Other transporters assist in the movement of these solutes out of the cell across its apical surface. As a result, solutes from the blood, particularly chloride, are secreted into the lumen of these exocrine glands, increasing the solute concentration within the lumen and causing water to be secreted out through osmosis. | |||||||||||||||||||||||||||||||||
| 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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4.7 ★★★★★
Based on 1454 reviews
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Product Reviews
★★★★★ 5
Excellent value, consistent fit, cool, comfortable and durable.
Size: Large, Color: Army Green
I have purchased 20 plus of these shirts in the past six months. The first batch in spring, figuring if any inconsistencies or quality issues arose upon delivery, no problem. Amazon's return policy has always been hassle free. After trying on the fourth shirt and noticing no outstanding flaws or differences, all were simply opened, laundered and hung on a hanger. The material is 100 percent cotton, but assuming they are removed from the dryer immediately (medium heat) - no ironing necessary. No appreciable shrinking is observed and no outliers in the batch.
My sport shirt size is men's US large regular fit. Other relevant sizes - 17.5 inch neck, 46 inch shoulders, 34 inch sleeves and 36 inch waist. Overall fit of these shirts is of appropriate roominess without bagginess. Shirt tail length is spot on, work dress code requires tucked in, and they stay tucked in. Casual requires not tucked in, and unless I am reaching for the sky with both hands while on tip toes, the shirt tail/ belt line is not breached. The sleeve length could have been extended by an inch since they taper to an elastic cuff, requiring that the sleeves are pulled back to original position following an extended reach. But given the choice, the sleeves as is are preferred over a straight cut without the elastic cuff.
As an upgrade to the T shirt appearance of the shirt tail when not tucked in, the side seams end with a reinforced, one inch inverted V, a feature not typical in this price range.
To date, durability has not been an issue. Fraying, fabric pilling and fading have not occurred. It is quite difficult, if not impossible, to differentiate the age of two duplicate color shirts purchased in two separate batches.
Again, excellent value, consistent fit, cool, comfortable and durable!
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Reviewed in the United States on October 30, 2017
★★★★★ 5
Great shirt.
Size: XX-Large, Color: Coral Orange
Purchased to replace an older coral polo shirt. Absolutely love the color and texture.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 7, 2026
★★★★★ 3
Sometimes Good, Sometimes Not So Good...
Size: X-Large, Color: Aqua
I've bought several of these shirts in the past, and I've always been very happy with them, right up until my latest purchase. Apparently, depending on which color/style you choose, the shirts may be manufactured in a completely different country. And despite Amazon's efforts to make sure their clothing brands have consistent sizing across the board, it's now clear to me that some of those countries are not on the same page. Case in point: Guatemala.
My latest Amazon Essentials polo shirt purchases consisted of the colors 'light yellow' and 'rust'. Now, the light yellow shirt seems fine. Maybe a tiny bit smaller than my previous shirt purchases, but nothing major. (I think it says made in Chihuahua - but it's hard to tell.) However, the rust colored shirt is drastically smaller than all my similar-sized Amazon polo shirts. I bought it in size Large, but the shirt tail alone is nearly four inches shorter than my previously purchased Amazon Essentials polo shirts. And while the Large-sized Amazon polo shirts normally fit me quite comfortably, with a little bit of room to spare, this rust-colored shirt is skin-tight. If I didn't know any better, I'd swear it was a size Medium masquerading as Large.
All of my previous shirt purchases (I now realize) are labeled as being made in either Madagascar or Vietnam. Those are great. They fit wonderfully. But this new rust-colored shirt (once again, same brand - Amazon Essentials) is labeled as made in Guatemala. And let me tell you, the factory in Guatemala is not on the same page as the factory in Madagascar. Not even close.
So yeah, these are great shirts. So long as you happen to buy the colors/styles that are being made in Madagascar or Vietnam. But buyer beware if you happen to get one from the factory in Guatemala, because those guys are using a whole different set of rulers over there. And unfortunately, there's no way to tell until it arrives on your doorstep.
I'm normally a big fan of Amazon brand products. But this experience has made me quite a bit more wary about buying their clothing items in the future. It really is so very important that their brands have consistent sizing across all colors and styles.
P.S. I actually bought the rust-colored shirt TWICE, because I at first assumed I must have gotten a mislabeled product. But, nope. Both rust-colored shirts are the same, drastically-smaller size, and both are labeled made in Guatemala.
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Reviewed in the United States on June 9, 2020
★★★★★ 5
Nice, well fitting and inexpensive Polo.
Size: Large, Color: Red
Don't be deceived by the low price. This is a nice shirt. To me the shirts are a bit "oversized" which I found to be perfect. The large is a "roomy" size large which is what I like. I previously ordered an XL which is my normal size in polo shirts and it was too big, and hung way low. Easy return of the XL and fast shipping of the L was appreciated. Going on a cruise, so ordering a few collared shirts. This is a keeper.
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Reviewed in the United States on January 23, 2026
★★★★★ 4
Decent, definitely got the job done
Size: X-Large, Color: Light Grey Heather, Size: X-Large, Color: Light Grey Heather
The came out to be a pretty decent quality for the price. Was going to a polo match and just needed something to creat on for my date
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Reviewed in the United States on April 1, 2026
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