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Description
Human FOXP3 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 utilizes 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 Forkhead Box Protein P3 (FOXP3). 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 Forkhead Box Protein P3 (FOXP3) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Forkhead Box Protein P3 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | FOXP3 (forkhead box P3), also known as scurfin, is a protein involved in immune system responses. As a member of the FOX protein family, FOXP3 appears to play a master role in regulating pathways involved in the development and function of regulatory T cells. In cancer, excessive regulatory T cell activity can prevent the immune system from destroying cancer cells. In autoimmune diseases, insufficient regulatory T cell activity allows other autoimmune cells to attack the body's own tissues. While the precise mechanisms of control remain unknown, FOX proteins belong to the forkhead/winged-helix family of transcriptional regulators and are hypothesized to exert control during transcription through similar DNA-binding interactions. | |||||||||||||||||||||||||||||||||
| 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 | 0.31-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates, cell lysates, and other biological fluids |
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4.6 ★★★★★
Based on 294 reviews
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Product Reviews
★★★★★ 3
Way too narrow
Size: 9.5 Wide, Color: Burgundy Polished Full Grain
I ordered a wide in this shoe, but it was still way too narrow. I had to return them. Otherwise, it looked like a nice quality shoe.
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Reviewed in the United States on February 12, 2026
★★★★★ 5
I usually wear 11.5's, but I buy 1/2 size smaller to accommodate for stretch. Leather relaxes.
Size: 11, Color: Burgundy Polished Full Grain
5 STARS: These shoes are awesome. I keep coming back for more. I have major foot problems, have tried all kinds of shoes, orthotics, and inserts... but believe it or not, these shoes work better than any of that stuff. I wear them daily. I buy lots of them, for home, for church, and I'm getting my first pair of J&M boots today, for winters in Indiana... Brrrr
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on December 18, 2025
★★★★★ 5
Stop complaining about the size and review the actual product! (Rant, Review, & Recipe)
Flavor Name: Culinary, Size: 1.06 oz (Pack of 1), Flavor Name: Culinary, Size: 1.06 oz (Pack of 1)
I was a little hesitant at first to buy this starter size pouch because of the price. I wanted good quality but kept asking myself, "What if I don't like it?" The company offers a 100% refund on their product so I went for it. Problem solved!
RANT: The biggest complaint I see from people on here is in regards to the amount you are getting for the price. They are not reviewing the quality of the item within the packaging or the flavor, etc. If you don't review the actual product and focus only on the quantity or packaging, the review is mostly worthless. As consumers, we get enough worthless reviews that are paid for, we don't need more worthless reviews because people only want to focus on the quantity and packaging and not review the quality. All are important in a review but the quality should be what is focused on.
Why is that that because it is a smaller size, people feel entitled to receive it for next to nothing? These are probably the same people that pay almost $2 for a 20 oz bottle of soda while the 2 liter bottle is only $1 and they don't bat an eyelash.
What they fail to understand is that making smaller sizes like this is not very cost effective for them to make. It probably costs them the same, or close to the same amount of money to purchase the bags they use for this size as it does for a larger size. These sampler sizes don't sell as well either. Unlike the bottle of soda that is "personalized" in size for the consumer to grab and go, this is a product that takes time and effort for the consumer to prepare. It is not meant to be a personalized size. If it were, people would have no problem paying for the convenience. Consumers are so spoiled and don't understand the costs associated with what it takes to import, pay the farmers a fair wage, the manpower and materials it takes to package the product, etc. I can guarantee you however, if a fair wage is not paid, someone is bound to call the 5:00 news! If you want sample sizes, fair wages, and the like, the consumer is going to have to pay for them or the company will go out of business really fast.
How many people have purchased an actual vanilla bean? Or saffron? Or truffles? No one seems to have a problem paying a high price for these items but complain about $10 for 30g of quality culinary matcha. Just like those items, a little matcha goes a long way.
It depends on how you plan to use it. If you are going to bake with it, buying a starter size is not cost effective for you.
If you are using it to make a tea latte, which is why I bought it, this is still gives you enough for 15 lattes that are 16 oz in size making them roughly $1.63 each (my recipe for this is below). Costs will vary depending on your ingredients of choice, and the area you live in. My numbers are based on $10 for 15 servings of this matcha and $0.08 per oz of organic vanilla almond milk in Austin, TX. Go to a coffee shop and you will pay at least $5 for a matcha latte of the same size, which is yet another thing buyers won't complain about. OK, RANT OVER...ON TO THE REVIEW!
REVIEW/COMPARISON: I used to hate the taste of Matcha but after years of taste bud readjustment, I decided to give it another try at Starbucks. They use Teavana brand for their lattes which is like Jade Leaf and does not use fillers and is organic. I try to avoid dairy so I used almond milk for my iced latte instead. Neither their tea or almond milk is sweetened so I added 3 pumps of vanilla to their venti size (they usually use 6 if you want it sweetened). Something about their almond milk leaves a bad aftertaste but before that hit, it was really good. Like most people, I can't afford $5 a day for one of these so I decided to buy the powder to make on my own at home (I was a barista in a former life so I often do this with my coffee concoctions too).
What I don't like about the Teavana brand is that it is much more expensive ($25 for 40g) and they claim it is ceremonial grade and yet, it looks and tastes exactly like Jade Leaf's culinary grade. In the brief research I did before buying, not a single self-proclaimed matcha aficionado EVER said to use ceremonial grade for a latte. They all very plainly stated that it was NOT for culinary use and that the taste is quite different. It is also higher in chlorophyll so it is a much brighter green color. That begs the question as to why Teavana, which looks and tastes the same as Jade Leaf is labeled ceremonial grade and being used for lattes when it is clearly the same culinary grade.
This morning, I made my first iced latte with this stuff from Jade Leaf. Using an organic, vanilla (sweetened) almond milk I found it was perfectly sweet and still had the very slight hint of the bitterness that comes from green tea. No weird almond milk aftertaste like the one from Starbucks. Needless to say, in the end, I will be buying this again in a larger size. It does say to store it in the fridge so that is where mine went when I was done with it. I plan to do a juice fast soon and a few recipes I have seen call for green tea for an extra metabolic and antioxidant boost. I plan to use this instead since it is so much more nutritionally dense than regular tea leaves.
RECIPE: They offer some recipe options on their Amazon listing and also on their website, but this was my super simple one that I threw together and was delicious.
Iced Matcha Vanilla Almond Latte:
4 oz (1/2 c) hot water (not boiling but barely steaming to help dissolve the matcha easily)
1 tsp Jade Leaf Organic Matcha
12 oz (1-1/2 c) cold vanilla flavored almond milk, or other milk of choice
Sweetener of choice, to taste (optional, I don't use any because my almond milk is already sweetened)
Ice cubes
NOTE: Sweetening options will have to be done at different steps depending on what you are using. If you are using liquid sugars like Torani or homemade simple syrups, those can go in at any time. Honey, agave, coconut nectar, sugar, and artificial sweeteners, should be blended in when you whisk the matcha into the hot water otherwise they will not dissolve properly and will settle at the bottom of the cup.
1. In a glass measuring cup, add 4 oz hot water.
2. Add 1 tsp of matcha powder and whisk together (see note for adding optional sweeteners)
3. Pour into a 16-20 oz cup over a few ice cubes to cool it.
4. Top off with the 12 oz vanilla almond milk (or other milk of choice) and stir to blend.
Add more ice if desired.
ENJOY Y'ALL!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on July 24, 2017
★★★★★ 5
Matcha Powder
Flavor Name: Culinary, Size: 1.06 oz (Pack of 1)
Jade Leaf Matcha has become my go-to matcha powder, and I keep coming back to it time and time again. The quality is consistently great, with a fresh flavor and vibrant green color that mixes well into drinks and recipes. I use it regularly for matcha lattes, smoothies, and even baking, and it always delivers great results.
For a culinary-grade matcha, I think the flavor is excellent—not overly bitter and smooth enough to enjoy daily. It blends easily and provides the perfect matcha taste without breaking the budget. I’ve tried other brands over the years, but this is the one I continue to repurchase. If you’re looking for a reliable, high-quality matcha for everyday use, I highly recommend giving this one a try.
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Reviewed in the United States on June 7, 2026
★★★★★ 5
Good taste,
Flavor Name: Culinary, Size: 3.53 oz (Pack of 1)
This matcha has a nice, rich flavor and works really well for lattes. It mixes easily without clumps and the color is vibrant. Good quality for the price and perfect for daily use.
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Reviewed in the United States on April 26, 2026
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