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Description
Human FAM3D 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 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 sequence similar family 3 member D (FAM3D). 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 sequence similar family 3 member D (FAM3D) in the sample. The absorbance (OD value) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Sequence similar family 3 member D ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Family of sequence similarity 3, member D (FAM3D), a member of the FAM3 family of cytokines, possesses a four-helix bundle structure. 11 FAM3D is constitutively expressed in the gastrointestinal tract, and its expression and production are regulated by nutritional factors. Specifically, FAM3D is upregulated in humans after consuming a high-fat diet. FAM3D upregulates and activates Mac-1 (macrophage antigen-1) in neutrophils, and inhibition of FPR1 (formyl peptide receptor 1) or FPR2 significantly blocks FAM3D-induced Mac-1 activation, indicating that the effects of FAM3D are dependent on these two FPRs. | |||||||||||||||||||||||||||||||||
| 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 | Tissue homogenates, cell lysates, and other biological fluids |
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4.8 ★★★★★
Based on 987 reviews
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Product Reviews
★★★★★ 5
Sleep-aid
I've used Oracoat XliMelts for years at night time. I usually only need one for overnight. Keeps my throat dry so I don't keep waking up with a dry mouth.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on February 8, 2026
★★★★★ 5
Finally A Product that Works For Me
I have severe dry mouth. It's due to medication which I need. It was so bad my tongue would stick to the roof of my mouth and I couldn't open it again. I had really bad gun problems and lots of dental work from the infections that developed. Eventually I had to give Up a valuable medication. That was the last century and toothpaste and mouthwash weren't enough. I am.now taking medication that I wouldn't be able to if it weren't for Xylimelts. I found keeping up with the right mouthwash and using this does the trick. I do need some squirts of Biotene spray. That's instant saliva. Keeping this pouches against the gum has been a game changer. It doesn't show. I don't feel it except when it runs out, gets tiny and crumbles into my mouth. That is about two hours. A friend used it and it lasts four hours. I dont remember if I felt it after the first couple of days. It's curved design makes it undetectable. I think the first day of two I kept checking it stayed in place. It does. It doesn't need any particular placement. I put It on the tip of my finger and wherever it goes is fine. This really works. Finally a product that does work for very dry mouth. At night I put two in and they're still there in the morning. I m a mouth breather so the dry mouth was really hard on me comfort-wise. People try to be kind and suggest you have a drink of water. They don't know it's not thirst. I will say taking sips of whatever you like helps. But in the last century I had to give up a valuable medication because my tongue stuck to the roof of my mouth so badly I couldn't open it again. Really embarrassing. It wasn't healthy for my gums either. This does it for me. I hope this helps other people who suffer from bad dry mouth or need it less often
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Reviewed in the United States on July 17, 2019
★★★★★ 5
I can Sleep Again!
I can get terrible dry mouth while sleeping. These have made it so I can sleep the whole night with out waking up to drink water. I love theses. Gave me back a good nights sleep. I use Two every night. Simple get it a little wet, then I place gently and hold to the outside of the top left molar for a few seconds. The glue will hold it in place with out fear of swallowing. Then I put one on the top right side. They stick well to teeth and can be easily removed. If you leave it alone and fall asleep, it will stay in place for 4-6 hours, providing gentle lubrication to keep your mouth moistened while sleeping. They make your mouth water a little, and the xylitol gives a little lubrication. These are great for mouth breathers and people with sinus issues. These are a real game changer for the better.
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Reviewed in the United States on May 25, 2019
★★★★★ 4
Xylimelts
Have been using for years. So glad they have these without the mint. Just wish they lasted longer at night but they do the job. Taste is pleasant and they are easy to use.
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Reviewed in the United States on December 16, 2025
★★★★★ 5
Dry Mouth Relief
Having been using these for quite awhile now for a severe dry mouth. They really help to make the mouth create saliva, thus protecting the teeth from erosion, etc. They are discs with a dark side which is placed above the teeth on the gum. It will stick there in a minute or two and remains in place until it melts and disappears. I use two at night and have never had a problem with them coming loose, this creating a choking hazard. They remain i place and melt. Sometimes I wake at night and my mouth is dry, so put a couple more in and I'm good to go until morning. I would recommend these to anyone with a dry mouth. There is no bad taste. They are somewhat expensive when using long term, but worth it to me. Have not found them locally, although Walmart has them online...same price or a little more than Amazon. Will continue to buy from these sellers.
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Reviewed in the United States on June 21, 2017
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