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Description
Mouse TIMD4 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 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 capture antibody against T-cell immunoglobulin and mucin domain containing 4 (TIMD4). 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 T-cell immunoglobulin and mucin domain containing 4 (TIMD4) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse T-cell immunoglobulin and mucin domain containing 4 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | T-cell immunoglobulin mucin domain-containing protein 4 (TIMD4), also known as T-cell membrane protein 4 (TIM-4), is a protein encoded by the TIMD4 gene. It contains an IgV domain with an integrin-binding site and a unique metal-ion-dependent ligand-binding site for phosphatidylserine. It also contains a mucin domain with high levels of O-glycosylation. Unlike other TIM proteins, it lacks a tyrosine phosphorylation motif in its intracellular tail domain. Unlike other TIMs, which are primarily expressed on T cells, it is expressed on APCs, such as dendritic cells and macrophages. It serves as a ligand for TIM-1 and also as a receptor for phosphatidylserine. However, its phosphatidylserine binding does not mediate signaling, but rather functions more as a tethering receptor. It can also inhibit naive T cells through binding to non-TIM-1 receptors, but once T cells become active, it acts as an active regulator, helping to maintain their activity. TIM-4 expression on macrophages plays an important role in maintaining their homeostasis. It plays an important role in Th2 development. Therefore, it plays a role in the development of allergies. It is also considered a factor in tumor development. It also mediates autophagy at tumor sites, which leads to reduced antigen presentation and increased immune system tolerance to tumors. | |||||||||||||||||||||||||||||||||
| 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 | Tissue homogenates, cell lysates, and other biological fluids |
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4.1 ★★★★★
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Product Reviews
★★★★★ 5
Small Valet Tray Nightstand Organizer (Black)
Color: Black, Color: Black
I really love the leather look of this valet tray. It’s both stylish and functional, giving a classy touch to my nightstand while keeping small items neatly organized. The PU leather finish feels smooth, looks high-end, and the construction feels sturdy.
Pros:
Elegant PU leather design with a textured finish
Compact 8x8 inch size fits perfectly on nightstands, dressers, or desks
Multi-purpose — great for keys, watches, jewelry, remotes, or glasses
Water-resistant surface makes it easy to wipe clean
Adds a decorative touch while being practical
Makes a thoughtful and stylish gift
Cons:
The sides are a little low, so bulkier items may slide out
Only available in limited colors compared to some other organizers
Overall, this is a really nice valet tray. It looks more expensive than it is, keeps my essentials in one place, and adds a clean, organized look to my space. Definitely worth it if you like the leather aesthetic!
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Reviewed in the United States on September 20, 2025
★★★★★ 5
Fits many trinkets and looks good
Color: Black
Simple little organizer for trinkets. Easy to snap together into a box shape and holds keys, pens, vapes, and other bits and pieces. My partner is happy I ordered it!
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Reviewed in the United States on October 4, 2025
★★★★★ 4
Excellent one
Color: Black, Color: Black
Build
These valet trays are perfect for placements on nightstands, sofas, end tables, etc. They are very compact and unobtrusive. Their size is ideal for holding daily essentials without taking up excessive space, and blending seamlessly with various decor styles.
Material
Constructed with a soft and durable synthetic surface, the tray is gentle to the touch and provides a wipeable and easy to clearn finish. The black color complements any decor and the materials prevent damage to surfaces they are put on.
Functionality
These trays serve as excellent catch-alls for remotes, phones, earbuds, glasses, jewelry, watches and other small items, helping keep sofas, nightstands and tables organized and clutter-free. The sturdy, level construction ensures stability, and the trays hold essentials like phones, TV remotes, and personal accessories effectively.
Value for Money (VFM)
Given their functionality, design, and affordability, these valet trays offer good value. They enhance organization without compromising style and are suitable for various rooms in the home. Will definitely recommend them.
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Reviewed in the United States on September 30, 2025
★★★★★ 5
Great Tray For Storing My Everyday Carries
Size: 8.5" x 5.8" x 1.9", Color: Black
My everyday carry items (wallet, keys, etc.), used to just get thrown around wherever when I'd get home, but this little tray makes a great, defined, space for me to keep those things when I don't need them on me. The soft lining reduces the obnoxious sound of throwing keys on a hard surface, and of course keeps me from scratching up a surface. The build feels quality and I haven't seen any damage to it yet. Also the all black version with gold buckle looks quite good and adds a subtle touch of luxury. Absolute recommend for the price.
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Reviewed in the United States on April 15, 2026
★★★★★ 5
Just the right size to empty your pockets.
Size: 8.5" x 5.8" x 1.9", Color: Orange
Perfect size to empty my pockets when I get home. Fits my wallet, keys, AirPods, etc. excellent quality.
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Reviewed in the United States on March 21, 2026