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Description
Mouse TIM-3 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. Preparation of standard gradient working solution: Add 1mL of universal diluent to the lyophilized standard, let it stand for 15 minutes to completely dissolve, then gently mix (concentration is 2000pg/mL). Then dilute to the following concentrations: 2000pg/mL, 1000pg/mL, 500pg/mL, 250pg/mL, 125pg/mL, 62.5pg/mL, 31.25pg/mL, and 0pg/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each tube. Pipette 500uL of the 2000pg/mL standard working solution into the first EP tube and mix thoroughly to make a 1000pg/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube is used 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 T-cell immunoglobulin mucin receptor 3 (TIM-3) 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 T-cell immunoglobulin mucin receptor 3 (TIM-3) 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 mucin receptor 3 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | T-cell immunoglobulin and mucin domain-containing 3 (TIM-3), also known as hepatitis A virus cellular receptor 2 (HAVCR2), is a protein encoded by the TIM3 gene. TIM-3 was first described in 2002 as a cell surface molecule expressed on IFNγ-producing CD4+ Th1 and CD8+ Tc1 cells. Subsequently, its expression was detected on Th17 cells, regulatory T cells, and innate immune cells (dendritic cells, NK cells, and monocytes). | |||||||||||||||||||||||||||||||||
| 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 | 31.2-2000 pg/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates, cell lysates, and other biological fluids |
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4.7 ★★★★★
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Product Reviews
★★★★★ 5
Liquid facial magic!
Scent: Coconut Water, Size: 12 Fl Oz (Pack of 1)
I love this stuff. I use it daily as part of my facial routine and it has become one of those products I don’t even think about anymore because it just works.
It smells good without being overpowering and does a great job of helping control excessive oil buildup on my face. Nothing harsh, nothing drying, just clean and refreshed skin. Right after using this, I follow up with a facial moisturizer and the combo works beautifully. My skin feels balanced instead of tight or angry, which is exactly what I’m going for.
I know the label says men can use this as an aftershave, but that’s a hard pass for me. I stick to soothing, purpose-built post-shave products, specifically from Wholly Kaw. That stuff smells like it was handcrafted by skincare deities who truly care about your face. This toner is staying firmly in the “daily skincare” lane, where it shines.
Another big plus is how gentle it is. No burning, no weird tingling, no regret. Just a calm, refreshing step that makes my skin feel like it’s being treated with respect.
And the price? Under $10. That alone keeps me coming back. Honestly, I think they could charge a few dollars more for how well this performs, but I won’t complain about paying less for fantastic facial liquid magic.
Simple, effective, affordable, and a permanent part of my routine. My face approves.
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Reviewed in the United States on December 21, 2025
★★★★★ 5
THAYERS Lavender Witch Hazel Toner: Time-Tested Elegance in a Bottle!
Scent: Lavender, Size: 12 Fl Oz (Pack of 1)
Embarking on a journey down memory lane with THAYERS Lavender Witch Hazel Facial Toner has been nothing short of a delightful reunion. This timeless brand, known since my youth, has proven its mettle yet again. Let's dive into the aromatic, moisturizing wonder that is the Lavender Witch Hazel Toner:
Quality Infused with Elegance (5/5): THAYERS brings a touch of timeless elegance to skincare, and this lavender-infused toner is no exception. The quality is unmistakable, leaving my skin feeling pampered and rejuvenated after each use.
Cleansliness at its Best (5/5): The toner's prowess in cleansing is unparalleled. It effortlessly removes dirt and impurities, unveiling a refreshed and squeaky-clean complexion. My skin thanks me for this daily ritual of cleanliness.
Moisturizing Magic (5/5): The moisturizing effect is like a gentle caress for the skin. Unlike some toners that leave your face feeling parched, THAYERS Lavender Toner ensures that my skin is not just clean but incredibly moisturized after every application.
Refreshing Elixir (5/5): Ah, the refreshing burst of lavender! While opinions on the strength of the scent may vary, I find the lavender notes to be a delightful touch. The toner rejuvenates not just my skin but also my senses, making each application a mini spa moment.
Better than the Expensive Competitors (5/5): Sometimes the best things come in unassuming packages. Users have attested that this toner outshines even pricier competitors, proving that quality doesn't always have to break the bank.
Mixed Durability Verdict (4/5): Opinions are divided on durability, but for me, the overall benefits outweigh any minor considerations. The toner's efficacy in daily use makes it a staple in my skincare routine.
Generations-Long Trust (5/5): THAYERS is a brand that has stood the test of time, transcending generations. Its legacy of trustworthiness extends beyond my own experience, connecting with others who have relied on its skincare wonders.
Lavender Scent Aspiration (4/5): While I had hoped for a stronger lavender scent, the subtle notes are still a pleasant addition to my skincare ritual. The product's efficacy compensates for any scent expectations.
In conclusion, THAYERS Lavender Witch Hazel Facial Toner is a testament to the enduring charm of a trusted brand. Its quality, cleanliness, and moisturizing benefits make it a staple in my skincare routine and a connection to the past. Whether you're a longtime fan or a newcomer, this toner is a timeless treasure for your skin. Cheers to THAYERS for continuing to deliver skincare excellence!
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Reviewed in the United States on February 17, 2024
★★★★★ 5
Lasts awhile! Good product
Scent: Rose Petal, Size: 12 Fl Oz (Pack of 1)
Cleans without overdrying. Leave skin feeling clean, no stickiness. Pleasant light scent that doesn't linger. I get my moister from other products so this one idk if it moisturizes by itself. Good value for the money, you get a lot so it lasts awhile.
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Reviewed in the United States on May 10, 2026
★★★★★ 4
Love! Smell is nice, helps with dry skin
Scent: Cucumber, Size: 12 Fl Oz (Pack of 1)
I have been using Thayers for years. My skin is sooo dry and tight post shower. I put this in a small mister bottle, and Mist my face.
Scalp- my scalp gets itchy, I use this, witch hazel, apple Cider Vinegar and water in a dropper. Works amazing!
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Reviewed in the United States on March 24, 2026
★★★★★ 5
Great rose toner!
Scent: Rose Petal, Size: 12 Fl Oz (Pack of 1), Scent: Rose Petal, Size: 12 Fl Oz (Pack of 1)
As an aesthetician, this is my go to over the counter toner. Rose is great for your skin and the ingredients in this are great. I am extremely sensitive, and this does not irritate my skin at all. It keeps it soft and clear.
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Reviewed in the United States on May 6, 2026