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Description
Human SEMA4B ELISA KitProduct Specification Usage Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High precision pipette and gun tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37 constant temperature box 4. Distilled water or deionized water Sample processing and requirements: Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4 overnight, then centrifuge at 1000g for 20
Product Specification
| Usage |
Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High-precision pipette and gun tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37℃ constant temperature box 4. Distilled water or deionized water Sample processing and requirements: Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4℃ overnight, then centrifuge at 1000×g for 20 minutes, and take the supernatant, or store the supernatant at -20℃ or -80℃, but avoid repeated freezing and thawing. Plasma: Collect the specimen using EDTA or heparin as an anticoagulant. Centrifuge the specimen at 1000 × g for 15 minutes at 2-8°C within 30 minutes of collection. The supernatant can be assayed or stored at -20°C or -80°C, but avoid repeated freezing and thawing. 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 the tissue and mince it. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1 g of tissue sample to 9 mL 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 the supernatant can be assayed. Cell Lysis Buffer: Gently wash adherent cells with ice-cold PBS, then trypsinize and collect cells by centrifugation at 1000 × g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash the collected cells three times with ice-cold PBS and resuspend them 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 freeze-thaw cycles 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 semaphorin 4B (SEMA4B) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by peroxidase (HRP) catalysis and to yellow by acid. The intensity of the color is positively correlated with the amount of semaphorin 4B (SEMA4B) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Semaphorin 4B ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Semaphorin 4B (SEMA4B), also known as semaphorin-4B or semaphorin-C, is a protein encoded by the SEMA4B gene. Its Sema domain is an interacting protein of LNM35 and plays a key role in lung cancer invasion. It inhibits the invasion of non-small cell lung cancer (NSCLC) by inhibiting MMP9 activation in a PI3K-dependent manner. It suppresses NSCLC growth in vitro and in vivo. Its adaptation in NSCLC cells alters cell proliferation. | |||||||||||||||||||||||||||||||||
| 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 | 0.156-10 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenates, cell lysates and other biological fluids |
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4.7 ★★★★★
Based on 11 reviews
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Product Reviews
★★★★★ 5
Perfect quality and Price
Color: Green, Size: 5Qt
Fantastic Price extremely great quality very heavy duty heat distribution is perfect and so is the heat retention it's excellent unhappy with this and the price is amazing and the colors are so vivid. It really makes cooking even more fun. I purchased the green and I also have a blue and I am so happy with my Dutch ovens.
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Reviewed in the United States on June 9, 2026
★★★★★ 5
Great cast iron enameled pot
Color: Red, Size: 5Qt
I love this pot. The first cast iron pot I got was not enameled and was supposed to be pre-seasoned. I tried to make steaks for our anniversary and they stuck something awful. I couldn't see the char against the black and scrubbed forever trying to get it off. After that I only get enameled pots with white or cream interiors. When you sear food it might stick but you can scrape the brown stuff off the bottom of the pan with a flat edged plastic or wooden spoon or spatula and it adds flavor to sauces. The pans also wash so much easier. If something doesn't want to come off you can just soak it in hot soapy water for a few minutes and it scrubs off. With the white or cream interior you can tell when you get the pot clean. Most can't be put in the dishwasher but they are easy to wash by hand. I also learned to cook at a much lower temperature than I did with my stainless steel cookware. The cast iron holds the heat so much better that it can get too hot if you use any temp over medium high. I still haven't figured out how not to burn minced garlic when I follow recipes. They always say to let the oil get hot until it shimmers but the garlic spatters something awful and quickly burns. Finally I did it my own way and put it in the pan before the oil gets too hot. The cast iron pot is heavy and on a glass surface cook top like mine you can't scoot it because it can scratch the surface. The handles get very hot too so you have to use hot pads, preferably silicone ones. The pot came with cute little teepee shaped hot pads that didn't really fit the handles well but fit the lid fine. This size is great for baking sourdough bread. Most recipes have you preheat the pot for an hour before adding the dough to bake. The lid fits tight enough to hold in steam from the dough to help the loaf develop a nice crust on it. I line the pot with parchment paper and the dough doesn't stick. You can also make soup and stews in this pot. It is safe on the cook top or inside the oven. I think the quality is good, but I'm not an expert on that. I know my pot hasn't shown any cracks or chips but I treat it carefully not exposing it to high heat when it is cold or a cold surface when the pot is hot so the glass enamel isn't exposed to temperature shock to avoid any cracking risk. Red is my favorite color and the pot looks great displayed on my cook top.
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Reviewed in the United States on December 31, 2025
★★★★★ 4
Heavy Smallish Handle Wonderful Dutch Oven
Color: Red, Size: 5Qt
As many have noted this lidded Dutch Oven is extremely heavy. But it cooks and cleans up beautifully, is well made and attractive.
In my opinion the handles are relatively small for such a heavy pot, my one complaint. The mitts are cute but more decorative than truly protective as the handles are so small. Just be very careful picking up and moving the pot especially from the oven.
I chose the red color which looked vibrantly cheerful in my white kitchen. The 5 qt capacity works for modest to regular-sized portions without second helpings for 4-6 people. However, if cooking for more people, want larger portions, 2nd helpings or leftovers, go with a larger pot.
Overall, given its great construction, price, cooking performance, and attractiveness, this is a no-brainer purchase if you won’t mind relatively small handles and a heavy pot.
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Reviewed in the United States on January 24, 2026
★★★★★ 5
Perfect for sourdough
Size: 4.3 Qt, Color: Olive
Beautifully made. Great addition to my kitchen!
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Reviewed in the United States on June 6, 2026
★★★★★ 5
Heavy-Duty Dutch Oven That Easily Feeds a Big Family
Size: 6.0 Qt, Color: Clay, Size: 6.0 Qt, Color: Clay
I am extremely impressed with this Amazon Basics enameled cast iron Dutch oven. The first thing I noticed when I took it out of the box was how substantial and heavy it feels, which immediately gave me confidence in the quality. The clay color is beautiful and looks even better in person, and the enamel finish is smooth and well done. I have a large family, so I needed something with plenty of capacity, and this 6-quart size has been perfect for big batches of soup, chili, pot roast, and homemade pasta sauce. It heats evenly, retains heat exceptionally well, and keeps food warm for a long time after cooking. The lid fits securely and helps lock in moisture, which made a recent pot roast come out incredibly tender after several hours of slow cooking. The wide handles make it easy to move from the stovetop to the oven, and despite the weight, it feels very sturdy and well balanced. Packaging was excellent and everything arrived in great condition. Considering the quality, performance, and attractive design, this Dutch oven offers outstanding value for the price. If you're looking for a durable cast iron pot that can handle family-sized meals and perform like much more expensive brands, this one is an easy recommendation.
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Reviewed in the United States on June 2, 2026