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Description
Human TFPI2 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 handling and requirements: Plasma: Collect the specimen using EDTA or heparin as an anticoagulant. Centrifuge at 1000g for 15 minutes at 2 8C within 30 minutes of collection. Remove the supernatant for testing or store at
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 handling and requirements: Plasma: Collect the specimen using EDTA or heparin as an anticoagulant. Centrifuge at 1000×g for 15 minutes at 2-8°C within 30 minutes of collection. Remove the supernatant for testing or store at -20°C or -80°C, but avoid repeated freezing and thawing. Pre-test preparation: 1. Remove the test 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 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 seven 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 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 Tissue Factor Pathway Inhibitor 2 (TFPI2) 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 Tissue Factor Pathway Inhibitor 2 (TFPI2) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Tissue Factor Pathway Inhibitor 2 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Tissue factor pathway inhibitor 2 (TFPI-2) is a member of the Kunitz family of serine protease inhibitors and a structural homolog of tissue factor pathway inhibitor (TFPI). TFPI-2 inhibits the tissue factor/factor VIIa (TF/VIIa) complex and multiple serine proteases, including plasminogen activators, plasminogen activators, factor XIa, trypsin, and chymotrypsin. TFPI-2 functions to maintain tumor milieu stability, inhibiting tumor invasiveness and growth, as well as metastasis formation. It also induces apoptosis and inhibits angiogenesis, which may significantly contribute to tumor growth inhibition. | |||||||||||||||||||||||||||||||||
| 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 | plasma |
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4.2 ★★★★★
Based on 11 reviews
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Product Reviews
★★★★★ 5
USED FOR SINUS ISSUES WITH GREAT SUCCESS
Size: 60 Count (Pack of 1)
Finally my 3 months of sinus congestion and cough are going away. My ears are also draining slowly too. This + oil pulling is helping with tooth sensitivity. Get this!
I have struggled with sinus issues my entire life with brief breaks from problems. I have had easily over 80 antibiotic prescriptions in my life for ear infections, 7 ear surgeries for a non-cancerous tumor, sinus infections, strep, etc. Last year the sinus problems began again and I was becoming dependent on antibiotics (5 infections in 3 months). After researching and concluding the bacterial infections were secondary to a fungal infection, my doctor and I decided to use a Nystatin both orally and powdered form in my netipot, and this cured the problem. However I was misdiagnosed with lyme this summer and was on antibiotics for a month. I also got strep throat, leading to another week of antibiotics. The sinus problems returned. I would have gone back to that doctor for more nystatin, however, I moved so that primary care, who was amazing and listened to me and trusted my science background, was no longer an option for me to visit. So I began researching ways to overcome ENT problems naturally and this probiotic came up as a good choice for sinus and teeth issues (I have also been having severe tooth sensitivity). And I am happy to say it is working. Additionally I have been putting the juice of kimchi up my nostrils and in my ears. The latter resulting in my ears draining onto my pillows for the first few nights.
I do not recommend Nystatin or any other fungal killer for that matter. After using Nystatin, I starting developing other problems like random joint and muscle pains, and I believe that is because the fungus was moving out of my intestines into the blood stream to escape the Nystatin (it has low absorption into the blood). I don't even recommend natural fungal killers like oil of oregano, pau d'arcco, neem, etc. I had been using for months and my intestinal candida was not improving. I truly recommend probiotics because biological control is always superior to chemical control. There is less chance for resistance since the organisms evolved together over millenia. Also building up diversity in your microbiome is hugely important. Get a good low-histamine probiotic to help with intestinal candida overgrowth and a good oral probiotic.
If you think you have systemic candida, I recommend the book Killing Candida Sweetly. I have found it helpful. I have been attempting to follow the protocols, but even with my moderate adherence, it is working.
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Reviewed in the United States on December 10, 2017
★★★★★ 4
This stuff works
Size: 60 Count (Pack of 1)
This stuff works. I'm 51 years old and about 2 years ago I started noticing stains close to the gum line on my otherwise pearly smile and also thicker plaque. I asked the dentist what to do as I'm ridiculous about brushing & flossing already.
She said I should think about getting a water flosser so I did and use it 2x a day. I've gone through 3 of them in 2 yrs time as they don't seem to last long. Anyhow at my next check up my dentist commented on how well my teeth were doing and I told her I had taken her advice and water flossing. I mentioned to her that I had read a study that said the amount of plaque in your mouth is directly correlated to the amount of plaque in your heart so I was determined not to have plaque.
I got a staph infection in my nose after a cruise and had awful garbage in my nose and running into my throat so I tried this stuff to heal myself. Wouldn't you know the fuzzy sweaters/plaque on my teeth lessened over time using these tablets and I no longer need to use the water flosser. I think I have a chronic Candida problem and have tried everything under the sun to get rid of it and I think this stuff corrects the digestive issues which lead to plaque and sinus problems. I'm about to order my second bottle because my teeth have no stains and actually look better than with the water flosser. Also I can easily pack the tablets for a trip and couldnt do that with the bulky water flosser. As a side note the dark circles and bags are better under my eyes so I think these tablets are healing everything related to the candida, plaque on teeth and sinus pouches. I haven't added any other vitamins or herbs to my daily regimen except these.
Would have given 5 stars if a full 2 months supply (60) was in the bottle, I hate when they short you just so you have to reorder early.
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Reviewed in the United States on October 31, 2019
★★★★★ 5
Great oral probiotic!
Size: 30 Count (Pack of 1)
Great oral probiotic! Tastes very good and has the right strains for oral health. Brushing, flossing and scraping are all key... and adding in a good oral probiotic is like the icing on the cake.
Not too expensive either. If you are looking for a product like this, I can highly recommend NatureWise.
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Reviewed in the United States on December 8, 2025
★★★★★ 3
Didn't Hurt, Didn't Help Much Either.
Size: 60 Count (Pack of 1)
Recently I was looking for an oral probiotic to help with sinus, gums and tartar for both myself and some pets. As a result, I purchased both this product and Great Oral Health's probiotics. Even though this product did have some positive effect as regards sinus pressure, it took two tables three times a day to work. Also, I noticed that an earthy, clay-like smell to breath in both myself and my pets as a result of using them. It didn't cause any harm, only that it was slightly disconcerting to smell like dirt.
The product from Great Oral Health didn't take as long to produce results, and there was a visible reduction in tartar. Also, no odor.
All in all, nothing majorly wrong with the product, except that it doesn't have results noticeable enough to warrant buying again.
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Reviewed in the United States on July 19, 2018
★★★★★ 5
Same probiotic strains, but a subtle formulation update
Size: 60 Count (Pack of 1)
I’ve used NatureWise Oral Health Probiotics (WiseBiotics) for a while and recently noticed a clear difference between the older and newer version — especially in taste and texture.
After comparing the labels side-by-side, there has been a formulation change. The probiotic strains are still the same (including BLIS K12 and M18), but the newer version now lists Organic Inulin (90 mg) as an active ingredient instead of just a minor “other ingredient.”
This seems to explain the changes I noticed:
• The tablets feel more powdery/chalky
• The taste is slightly different (a bit less crisp mint, slightly sweeter/fiber-like)
• The dissolve/chew experience is not quite the same as before
Inulin is a prebiotic fiber, so this change may actually help feed the probiotics — which is a positive. But it definitely changes the user experience.
For me, it’s not necessarily a dealbreaker, but it’s noticeable enough that others should be aware, especially if you’re sensitive to texture or ingredients like inulin.
Bottom line:
Same probiotic strains, but a subtle formulation update that affects taste and texture. Still a good product — just not identical to the previous version.
Hope this helps someone else who thought they were imagining things 🙂
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Reviewed in the United States on April 8, 2026