Pay in installments of $46.03 with
,
and
Shipping Estimate
USA
- USA
- CAN
- USA
- CAN
Ships within 48 hours · Estimated delivery Aug 10 - Aug 15
Promo Codes Available:
For Your Every Summer RSVP, with Code: SUMMER15
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. |
|||||||||||||||||||||||||||||||||
| 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 |
|
|||||||||||||||||||||||||||||||||
| 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. |
|||||||||||||||||||||||||||||||||
| 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 |
Shipping Notes
- Free Standard Shipping on $100+ Orders to the USA.
- Except Preorder products are shipped in 48 hours.
- Delivery to the USA:
- Standard Shipping : 3-10 business days
- If time is of the essence, please consider selecting expedited delivery for faster service.
Exchange/Return Notes
- We offer a 30-day return/exchange service after receiving.
- Final sale items are not eligible for returns or exchanges.
- To process your return/exchange, please contact us at [email protected]
- Please click here for more details>>> Return & Exchange Policy
4.8 ★★★★★
Based on 19 reviews
Sort
Product Reviews
★★★★★ 4
Helped to improve combustion in 12 year old Mercruiser 3.0L engine
Color: Blue, Number of Items: 1
I used the whole bottle in 1/2 tank of questionable gasoline from used boat from dealership, with no idea what mixture of gas water and particulates was in the gas tank in this 12 year old boat.
The dealer started up the boat, and it was taking a minute to get the idle going. He mentioned that the gas was a few months old, but I am guessing that is was more like 6 months old. This is the TKS engine, self priming. I think some water / condensate was in the tank. Before taking out to first trip, I added bottle and 5 gallons fresh gas. That trip to the lake, I still has some issues getting it to start easily. Ran OK, not great that day, went home, changed cap and rotor and wires. Next trip, 1 week later, the 3.0L started great, ran great, I am not sure if it took some hours to fully mix and interact with the old gas, neutralize the water, and perform it's reactions, before it made existing gas more combustible, but that is what it seemed like to me.
***UPDATE**** - I had strange fuel issues the next season after using this product. I had to replace the fuel pump and many other fuel delivery components including the carburetor. Not sure if this contributed to the fuel system failing, the long term effects outweigh the short term gains.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on September 25, 2024
★★★★★ 5
good buy
Color: Blue, Number of Items: 1
good buy
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 31, 2026
★★★★★ 5
Had water in my gas
Color: Blue, Number of Items: 1
Made my 150 outboard quit bogging down during acceleration
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 25, 2026
★★★★★ 5
Works. . .
Color: Blue, Number of Items: 1
Left 10% ethanol gas in my 3500 watt inverter and forgot about it. After I got it started, it was suging and alsmost unusable even with fresh gas. Drained all the gas and used a measuring cup to get the fuel/additive ratio right and let the generator run wiuth a 1000 watt work light. AFter about 15 minutes, the surging stopped and engine smoothed out. Happened faster than I expected. Taught me a lesson about gas decomposition. Promised the genny I learned a lesson to use stabilizer. Easier than doing a carb rebuild.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 2, 2025
★★★★★ 5
Tried as a last resort... It worked!
Color: Blue, Number of Items: 1
My 3.0 MerCruiser stern drive was hard starting and would not hold at idle and ran really rough under load. I put 2 cans of S*Foam Marine through the tank and replaced the fuel filter and the spark plugs. With only a slight improvement I decided to bring it to my dealer. They replaced the distributer cap, rotor and plug wires and sent me on my way. The idle was good, but under load (headway speed) it ran really rough. Brought it back and they swapped out the carburetor and lake tested it. Back at home it ran great for a one-hour ride. A week later I tried to start it and the idle was bad again (engine shaking, erratic RPM). Almost dragged it back to the dealer but thought I would try this first. Per instructions I added this to about 15 gallons already in the boat and then topped it off with 5 gallons of fresh gas. Let it idle at about 1500 RPMs for 15 minutes and WOW, it's purring like a kitten. Next time, this will be my first resort!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on August 6, 2024
recommand products
MICHAEL'S Blood Pressure Factors - 180 Vegetarian Tablets | Fluid Balance Support | Kosher
107.10
KarmaRipple's Year of the Dragon Colorful Double Dragons Playing Bead Bronze Protection Pendant Necklace
33.00
KarmaRipple's Pearl Bead Waterdrop Calm Peace Pendant Necklace
33.00
LIFETIME Diosmin Flavonoid Complex – Circulation & Vein Health (60ct, 30 Serv)
49.78
Liquid Magnesium Oxide 400mg Drops – High Potency Mg Oxide for Bone, Muscle & Heart Health (60ml)
45.98