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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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4.1 ★★★★★
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Product Reviews
★★★★★ 5
Great privacy option for townhome deck!
Color: White, Size: 6 Panel, Color: White, Size: 6 Panel
Deck privacy! I wanted a way to add some privacy to my second floor townhome deck. It’s been holding up well and doesn’t look junky! I used zip ties on one end so I can open and close it as I wish. Great solution to living in close quarters to your neighbors! I purchased the six panel and it’s a perfect size
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 3, 2026
★★★★★ 4
Flimsy
Color: Beige, Size: 8 Panel
It’s thinner than what I thought and flimsy.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on March 2, 2026
★★★★★ 5
Great room divider
Color: Black, Size: 4 Panel-88'', Color: Black, Size: 4 Panel-88''
This is a room divider and would work great as one! I’m using it as a cover for my car port/garage so you can’t look in and see my messy garage as easily and for what I’m using it for it’s a little not quite as sturdy as I need it to but I’m sure it’s not meant to be used outside anyways so I don’t fault the product since I’m not using it as intended… with that said though…. It still covers my garage beautifully and really does exactly what a I need it to! I’m sure the wind may blow it over if it gets too crazy but I don’t see it breaking by any means. The frame is metal and drilled together using long screws. It’s very well built. The cloth for the panels is this waterproof type material that’s kind of like the gazebo roof feeling. So it’s easy to clean and it’s easy to wipe. You can put it straight or bend the panels and either way it still stands up. You don’t HAVE to bend it to make it stand or anything. You can literally use it straight across like I am in the pictures. Overall very happy with this product and it’s also very tall so it truly covers a lot (hides a lot lol). I’m 5’8 so you can see it’s taller than me by the pic I took
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 4, 2026
★★★★★ 3
OK. Drag to setup. Needs more stable feet
Color: Black, Size: 4 Panel-88''
I was glad to find these.
The benefits of having them in a recording setup is helping suppressing noise for a vocal booth.
It's a layer that surrounds the recording box. Nice!
(But away from audio hardware or DAW).
Here is the not so nice part:
The feet!
It's able to stand (as it needs to do this). But it cannot be bumped. If it does, it's coming down onto something.
I believe the feet units are not wide enough (or long enough). That makes for an easier tumble risk. So far, they haven't, but it's like being in the woods over leaves - it feels dangerous with every step! (That's actually true).
They are tall enough, wide enough, material OK. But the feet for the main assembly do not seem sturdy enough.
There are ways to help that.
I'm thinking a 2x4 to augment the base. Make them wider. It can be done. It was a drag to set these up, and I'm sure to modify them like i'd want them. But, they do come in at a much lower price that other similar dividers.
Consider all this because I've seen more solid dividers ( with padding) that would basically be the 1/2 cost of these. But i'd think they'd be easier to assemble, stand sturdier, and without that feeling of an impending fall (or fail).
This is my opinion. I hope it helps.
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Reviewed in the United States on May 29, 2026
★★★★★ 1
wrong size of metal components. incorrect items. non functioning
Color: Black, Size: 4 Panel-88''
The owner of this product sent me the incorrect pieces when I tried to install. It didn’t fit. The pieces were missing holes and once I connected the wrong pieces after knowing now they’re stuck and I can’t return them even though Amazon was kind enough to refund me. I just want to warn people this product is such a disappointment because it looks good. It’s good price, but you can’t even assemble the thing very disappointed.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 31, 2026
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