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Description
Mouse DEFa6 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: Serum, plasma, 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
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: Serum, plasma, 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 freeze-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 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 1000×g for 20 minutes, and collect the supernatant for analysis. Preparation for the Assay: 1. Remove the reagent kit from the refrigerator 10 minutes in advance and equilibrate to room temperature. 2. Prepare a gradient standard working solution: Add 1 mL of universal diluent to the lyophilized standard. Let stand for 15 minutes to completely dissolve, then gently mix (concentration 1000 pg/mL). Then dilute to the following concentrations: 1000 pg/mL, 500 pg/mL, 250 pg/mL, 125 pg/mL, 62.5 pg/mL, 31.25 pg/mL, 15.625 pg/mL, and 0 pg/mL. Serial dilution method: Take seven EP tubes and add 500 μL of universal diluent to each tube. Pipette 500 μL of the 1000 pg/mL standard working solution into the first EP tube and mix thoroughly to make a 500 pg/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 Defensin Alpha 6/Paneth Cell Specific (DEFa6) 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 Defensin Alpha 6/Paneth Cell Specific (DEFa6) 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 6; Paneth Cell Specific ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Defensin α6, Paneth cell (DEFA6) is a protein encoded by the DEFA6 gene. DEFA6 is expressed in Paneth cells of the ileum. α-defensins are a family of microbicidal and cytotoxic peptides that protect the host against bacteria and viruses. However, DEFA6 provides protection against invading enteric bacterial pathogens by self-assembling into cellulose and nanomembranes that surround and entangle bacteria. Several α-defensin genes, including DEFA6, are clustered on chromosome 8. | |||||||||||||||||||||||||||||||||
| 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 | 15.6-1000 pg/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate and other biological fluids |
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4.1 ★★★★★
Based on 27 reviews
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Product Reviews
★★★★★ 2
Not linen
These aren’t linen, these are microfiber shorts. Fabric feels very cheap.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on July 12, 2025
★★★★★ 5
True To Size
These shorts fit true to size and are so comfortable. They have a drawstring waist, so you can adjust them for the best fit and wide legs. They are 55% linen and 45% cotton so they keep you feeling cool. The classic colors go with nearly any other colors or designs, making them stretch your wardrobe potential and saving you money. They were extremely wrinkled upon arrival, so I used my fabric steamer, and within 5 min they looked perfect. I love these shorts and wear them all the time!
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Reviewed in the United States on May 27, 2025
★★★★★ 4
Cute 2-pack pair of shorts
Color: Black, White, Size: Medium, Color: Black, White, Size: Medium
Cute shorts that have pockets and fit true to size. I love that they come in a pack of two as well. The black ones are my favorite and I feel like the fabric is a little softer on the black pair. The white ones are a little thinner than I would like. I was wearing an olive green colored underwear underneath them in the pictures in my review and though you can’t see them much, I do feel like they were slightly visible. The white pair also had a lot of loose strings on them which made me think the quality might not be as good as with the black pair. They washed easily and I hung them up to dry them because they smelled kind of funny when they arrived.
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Reviewed in the United States on June 7, 2025
★★★★★ 4
Good value for 2 shorts
These come vacuum sealed and have crazy wrinkles... so be prepared to do some ironing. I ordered the Mediums as I am a 6-8. The black pair feels a bit smaller than the beige pair... with both feeling tighter than I like for comfort. The look nice on, but feel like it constricts my belly a bit too much. Overall a nice basic set of shorts, perhaps size up for a more comfortable fit.
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Reviewed in the United States on June 15, 2025
★★★★★ 5
Super soft and comfortable!
I haven't bought new underwear in forever. I used to wear thongs, but now find them incredibly uncomfortable. The last few times I've tried to buy new underwear, the sizing was either way off (I'm a medium, but gave up after having to return them for a large and then extra large) or the quality was horrible (two brands stretch waaaay out and they can fall down while wearing them (I have larger ass-ets, so they shouldn't start to slide down during use). I tried these after buying some bamboo pyjamas and I love them! They are incredibly soft, comfortable, and stretchy. I can use them all day, and they work well, whether I'm lounging on the couch or wearing them under my spandex shorts at the gym, they don't ride up or slide down. They are both comfortable and look sexy, while providing support in all the right places. I haven't had them too long, so I can't speak on the durability, but they appear to be of good quality and I plan to order more, this time without the band.
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Reviewed in the United States on November 13, 2025
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