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Description
Human NCF4 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 10 ng/mL). Then dilute to the following concentrations: 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 ng/mL, 0.15625 ng/mL, and 0 ng/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each. Pipette 500uL of the 10ng/mL standard working solution into the first EP tube and mix thoroughly to make a 5ng/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube serves as a blank well; there is no need to pipette liquid from the penultimate tube. See the figure below for details. 3. Preparation of biotinylated detection antibody working solution: Centrifuge the concentrated biotinylated antibody at 1000×g for 1 minute 15 minutes before use. Dilute the 100× concentrated biotinylated antibody to a 1× working concentration with universal diluent (e.g., 10uL concentrate + 990uL universal diluent). Prepare and use immediately. 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 Neutrophil cytosolic factor 4 (NCF4) 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 Neutrophil cytosolic factor 4 (NCF4) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Neutrophil cytosolic factor 4 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Neutrophil cytoplasmic factor 4 is a protein encoded by the NCF4 gene. This gene encodes a membrane-regulated component of the superoxide-producing phagocyte NADPH oxidase, a multicomponent enzyme system important for host defense. This protein is preferentially expressed in cells of the myeloid lineage. It primarily interacts with neutrophil cytoplasmic factor 2 (NCF2/p67-phox) and forms a complex with neutrophil cytoplasmic factor 1 (NCF1/p47-phox). The latter further interacts with the small G protein RAC1, translocating to the membrane upon cellular stimulation. This complex then activates aflatoxin b, the membrane-integrated catalytic core of the enzyme system. The PX domain of this protein can bind to the phospholipid product of PI kinases, suggesting a role in PI kinase-mediated signaling events. Phosphorylation of this protein has been found to negatively regulate enzyme activity. Alternatively spliced transcript variants encoding different isoforms have been observed. | |||||||||||||||||||||||||||||||||
| 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 | 0.15-10 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates, cell lysates, and other biological fluids |
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4.5 ★★★★★
Based on 1286 reviews
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Product Reviews
★★★★★ 4
Soft...but fluffy and tall
Color: White, Size: Standard (Pack of 2)
I've only had these pillows for about a week. They are a great quality pillow. The cover is soft and seems well made. My only complaint is that they are VERY full pillows. If you're tall with broader shoulders, you'll probably love them. If you're shorter with a smaller frame you will probably feel like your head is too high and your neck at a weird angle (my problem). There seemed to be mixed reviews about softness....I would consider these a nice medium pillow. Soft filling but firm enough that your head doesn't sink to the bed. Definitely NOT a down-like pillow, so if that's what you're looking for, this is not for you.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 10, 2026
★★★★★ 5
Finally! Comfy pillows for a😴good night sleep
Color: White, Size: Queen (Pack of 4)
Just wow!!
Looking for new pillows that are comfortable & hold their shape. These are it !
I love anything by utopia. And these pillows did not disappoint.
Great value. Feels like a high end pillow.
As a side sleeper these are a keeper.
Highly recommend. And ill be purchasing more!
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Reviewed in the United States on May 18, 2026
★★★★★ 5
Soft, fluffy and surprisingly comfortable
Color: White, Size: Queen (Pack of 2), Color: White, Size: Queen (Pack of 2)
I really liked these pillows. I tried to show different things in my photos because I wanted people to see the details: the design, how fluffy they are, and how full and thick they actually look. They are so full that the pillowcase even looks tight once you put it on, which I actually loved because they look luxurious and well-filled.
At first glance they look very thick and high, but once you lay down they feel soft and fluffy instead of hard. They adjust comfortably and feel very cozy whether you’re sleeping, relaxing, or even reading before bed.
I think they offer a really good balance because they feel supportive without being stiff. Of course, pillow preference is very personal—some people like lower pillows and others like higher ones—but for someone who likes fuller pillows that are still soft and comfortable, these are great.
They are spacious, cozy, and feel like a very good purchase overall. I honestly don’t have any complaints so far. Definitely happy with them.
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Reviewed in the United States on May 15, 2026
★★★★★ 5
Great quality pillows. Very difficult wrapping.
Color: White, Size: King (Pack of 2)
The Pillows are the real deal. Very fluffy but with good support. I was surprised by the weight of the box they came in, pretty heavy so you know there’s good quality pillows inside.
The one negative is that the pillows are packed in very very tight plastic because they come compressed. But the thing is, it’s really a chore to open them. You have to be careful not to cut through the plastic to the material, but the plastic is put on so incredibly tight that it’s even hard to get your finger underneath.
But I found a way to open them. Near the top, insert A very flat metal ruler as far in as you could get it. Then cut through the plastic that’s above the metal ruler. That prevents a sharp tool from damaging the pillows. As you get through the plastic above the ruler, keep pushing the ruler further down into the package. Believe me, this simplifies what could be a very difficult unpacking. I am very surprised that the company that makes them does not insert the type of opener that you find on boxes you receive in the mail or even packages that you get from the grocery store where you just pull a tab and that opens the package.
These are really good pillows, but you must be very careful about using a sharp object when opening the box and when trying to cut through the plastic. I am a strong man and have the tools needed, a very flat metal ruler, and a box cutter, knife, or razor. Good luck. !!
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Reviewed in the United States on May 5, 2026
★★★★★ 5
Good quality
Color: White, Size: King (Pack of 2), Color: White, Size: King (Pack of 2)
These are so comfortable. I don’t think I even have to break them in. Soft and firm at the same time
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Reviewed in the United States on May 24, 2026
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