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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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4.7 ★★★★★
Based on 29 reviews
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Product Reviews
★★★★★ 5
Good hobo bag
Color: Grey
I love a good hobo bag for its laid-back style and functionality. It’s spacious without feeling bulky, and the slouchy design gives it that effortless, everyday vibe. There was no odor,material is a good thickness, not sure on durability since I just started to use it. The bag was bigger than I expected.
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Reviewed in the United States on April 22, 2026
★★★★★ 2
Strong stinky odor
Color: Army Green
I cannot share the stink of the purse. It fills small rooms with its odor. The bag is okay, but needs to be washed….
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Reviewed in the United States on April 16, 2026
★★★★★ 5
Perfect size!
I love a good large hobo bag! This one has a great slouch to it and you can fit a ton in it! I've had everything from my iPad and a makeup bag plus yoga clothes and still had room for a water bottle too! The fabric is gorgeous and although I'm not always keen on logo-prints, I like that the bag's print is so understated and discreet.
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Reviewed in the United States on May 22, 2026
★★★★★ 5
Large, non zipping, sturdy, stylish
large cream MK purse
I’m giving my thoughts on this purse that I got recently and here is what I liked.
Size-
This is a large purse. It has one large main compartment that is secured with a gold, snapping clasp. It doesn’t come apart unless you push both sides of this clasp.
1 pocket inside for my phone or whatever I need quick access to.
2 vertical outside pockets with working zipper. The pockets run the height of the purse and are narrow. I can get a skinny water bottle in there.
Not Adjustable
This comes with a short shoulder strap. The strap is not removable. There are two D-rings (H-ring) on each side that hold the strap so if I want I can add a long strap. Really though, this is not the kind of purse that would look good with a long strap. There is plenty of room to put the purse on your shoulder and it feels very secure.
Look and feel
It looks and feels like leather to me but it says PU which is not leather. I’m fine with that. It is firm in a sturdy strong way, having structure and can stand up on a table. The material is the kind you can wipe with a damp cloth. This purse is not going to get stained unless it’s something like ink that stains practically everything. Mine has monogram NK but if you don’t look real close, I think it looks a lot like a designer purse with the monogram m and k.
Function
I tend to change purses a lot so I keep things in smaller bags. For example, I have small bags for makeup, medical etc and they fit neatly in here. There is not a designated place for cards and cash so you’d need a wallet or you could put it in the inside pocket. I use a medium sized wallet. My iPhone 14 with case fits in the inside pocket and hasn’t fallen out yet. The 2 outside zipper pockets are large enough to hold my phone and my wallet, one on each side.
I can’t think of anything I’d add or change right now. This is a very nice bag to carry everyday. The holding capacity is similar to the neverfull.
I’m happy I got this and can recommend it to you. It’s nice looking, makes good use of space, and for me, was a good deal.
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Reviewed in the United States on March 15, 2026
★★★★★ 1
👎🏻
Color: Beige
0 stars
Poor quality
Arrived torn
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Reviewed in the United States on May 25, 2026