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Description
Mouse DEFa5 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: 1. 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: 1. 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. 2. 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. 3. 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. 4. 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 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 capture antibody against Defensin Alpha 5, Paneth Cell Specific (DEFa5). 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 Defensin Alpha 5, Paneth Cell Specific (DEFa5) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Defensin Alpha 5, Paneth Cell Specific | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Defensin, alpha 5 (DEFA5), also known as human alpha defensin 5 (HD5), is a protein encoded by the DEFA5 gene, which is expressed in Paneth cells of the ileum. Defensins are a family of microbicidal and cytotoxic peptides (antimicrobial peptides/AMPs) that participate in host defense and help maintain a balanced gut microbiota. DEFA5 is the primary AMP that controls the composition of the gut microbiota by selectively killing bacterial pathogens while sparing commensals. Defensins are small cationic peptides linked by three intramolecular disulfide bridges. They contain six intramolecular cysteine residues, forming a specific, immutable pattern of disulfide bridges that protect them from proteolysis and maintain their function within the intestinal lumen. | |||||||||||||||||||||||||||||||||
| 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 | Serum, plasma, tissue homogenates and other biological fluids |
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4.4 ★★★★★
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Product Reviews
★★★★★ 5
Great for portability and Many uses
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I use my iPad for work!! This case is a fantastic addition - easy to use when I’m on the couch or in the Office. Stand is easily changed from vertical to horizontal - Velcro handle can change large or small. My stylus also attached to the back so everything is ready on the GO
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Reviewed in the United States on May 14, 2026
★★★★★ 5
A Tough Everyday Shield That Turns an iPad Into a Travel-Ready Tablet
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I’m very impressed with this protective iPad case. It has a strong, rugged design that makes the tablet feel much safer for everyday use, especially for kids, travel, work, or anyone who needs extra protection. The black design looks modern, durable, and professional without being too bulky in appearance.
What I like most is how complete and practical it is. The built-in stand makes it easy to watch videos, study, read, or use the iPad hands-free, and the hand strap adds extra comfort when carrying or holding it. The included shoulder strap is also a great bonus because it makes the case even more convenient for moving around.
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Reviewed in the United States on June 10, 2026
★★★★★ 5
Great low cost USB C to 4K HDMI splitter adaptor
Color: Black
The Anker USB C Hub 5-in-1 USBC to HDMI splitter with 4K display with one powered USB c and 2 USB A ports works flawlessly and is a perfect travel companion or simple and inexpensive desktop dock to connect a moderan USB C only laptop to a large monitor up to 4K resolution. It supports 4K at 60 Hz so not for high speed gaming but it is flicker free for daily work where running at 60 Hz provides a clean looking 4K display. The USB C and USB A ports work perfectly. I already have four of these in different port configuration but wanted the 4K extension for travel and this was the best one for quality and bang for the buck.
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Reviewed in the United States on May 4, 2026
★★★★★ 5
Compact hub that punches well above its size
Color: Black
This hub handles 4K HDMI output and three data ports cleanly without any noticeable lag or connection issues. The powered ports on both the USB-C and USB-A sides are a useful feature for running peripherals without draining the host device. Compact enough to leave plugged in permanently without getting in the way. Works reliably on MacBook and Dell machines as described. Good value for a well-built hub with a solid feature set.
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Reviewed in the United States on June 8, 2026
★★★★★ 5
Compact, Travel friendly, Works well with ROG ALLY
Color: White
This is a compact and well built adapter, and it is travel friendly. I use it for my ROG ALLY when I don’t want it docked and still need charging and other device inputs. It has just about enough ports for what I need, usb-c power delivery port for charging (ROG ALLY can still run at maximum performance using this with a 100W charging brick and a cable that allows 100W power pass through). I use the remaining USB-C port and two USB-A ports for external hard drives or gaming controller 2.4Ghz receiver. Transferring files between ROG ALLY and an external SSD with this adapter is fast, could reach above 380 mb/s. All the device inputs were as advertised, “plug and play”. The hdmi port also gives me an option to connect my ROG ALLY to TV when I don’t have the dock which takes up more space for traveling. One thing to note, you will have to use the USB-C port on your PC which provides more power (usually at the back and not on the front of the pc), if not plugged in to a power source. When I received my first order, I plugged it into the front USB-C port of my mini PC, just for my wired keyboard and mouse, the computer didn’t recognize it and couldn’t get any input from my keyboard and mouse. I returned the first one and thought it wouldn’t work as I expected. A few days later when I was using another Anker 7 in 1 USB-C adapter, I ran into the same issue but I know that adapter had been working well, then I realized I had to plug it into the port on the back of the PC. So I got this again and tried, it just worked like a charm. Hope they actually mentioned it on their product page so I wouldn’t have to return the first one. The cable length on this adapter is just right, not too long or too short when using with ROG ALLY (not hanging, since the cable is pretty sturdy, when I picked up the ROG ALLY, the adapter just comes along).
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Reviewed in the United States on August 18, 2025