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Description
Human FⅧ 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 Coagulation Factor VIII (FVIII) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB converts to blue under the catalysis of HRP and to yellow under the action of acid. The intensity of the color is positively correlated with the amount of Coagulation Factor VIII (FVIII) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Coagulation Factor VIII ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Factor VIII (FVIII) is an important blood clotting protein, also known as antihemophilic factor (AHF). Factor VIII is encoded by the F8 gene. Defects in this gene cause hemophilia A, a recessive X-linked blood clotting disorder. Factor VIII is produced in the sinusoidal cells of the liver and the endothelial cells outside the liver, distributed throughout the body. This protein circulates in the bloodstream in an inactive form, bound to another molecule called von Willebrand factor, until an injury occurs that damages blood vessels. | |||||||||||||||||||||||||||||||||
| 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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4.3 ★★★★★
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Product Reviews
★★★★★ 5
My favorite... And I'm picky.
Size: 3 Ounce (Pack of 1)
I've become a little obsessed with sunscreen over the last past 5 years. I have olive skin and developed melasma after my first pregnancy. I learned the hard way that chemical sunscreens don't help if you have melasma - and they actually make the dark spots even worse. Also the active ingredients in chemical sunscreens (like Oxybenzone and Octinoxate) are absorbed by your skin, and may cause allergies, hormonal imbalance and be toxic overtime. After learning that, I started wearing only mineral sunscreen (active ingredient is zinc and/or titanium dioxide). These are not absorbed and works by creating a physical barrier that reflects the sun (some studies suggest that some nanoparticles of zinc may be absorbed by your skin, so to be really, really safe you should look for non-nano particles).
After trying a few brands, I realized the sunscreens with only zinc in their formulas worked better than the blends with titanium dioxide, but this is just my personal preference I guess. And the higher the percentage of zinc, the better.
Most pure mineral sunscreens out there are 10-ish % zinc, SPF 30, 40 min water resistant - which is ok.
Thinksport goes beyond that: It's 20% non-nano zinc, SPF 50+, 80 min water resistant. It really can't get any better than that!
The lotion is not extremely thick when compared to other sunscreens I've tried and I find it pretty easy to apply. As every zinc sunscreen, it does leave a little fine white cast, but honestly Thinksport is not bad at all! It smells nice, lightly citrusy - and the smell vanishes soon. And, it's not overpriced.
Best I've tried so far.
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Reviewed in the United States on April 15, 2016
★★★★★ 3
Works but will Dry Out Your Skin
Size: 6 Ounce (Pack of 1)
I am a swim instructor at an outdoor pool and this stuff DOES work. I can apply it at the beginning of a 5 hour shift and while I have experienced light tanning it has protected against burning completely without reapplication.
The white cast is significant though I don't mind it aesthetically, I know its there for protection . It does rub off on things though so consider yourself warned.
Major Con: it is incredibly drying, and the soap needed to scrub it off is A LOT. That coupled with chlorine is a lot for my skin . Still on my sunscreen journey.
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Reviewed in the United States on May 4, 2026
★★★★★ 5
Protected Our Skin and Smells Good
Size: 6 Ounce (Pack of 1)
I love this stuff! My husband and I bought it as a last minute grab for our honeymoon and it was well worth it! I'm very sensitive to the sun AND to sunscreens, so I needed something with simple ingredients that would also protect my skin without breaking me out. This not only kept away the burning rash I get from the sun, but protected my husband and I from any sunburn at all. It worked not only in dry conditions, but it stayed on for hours after we swam in the ocean. We also liked the fact that it smelled herbal, almost like pizza/Italian seasoning. That didn't bother us because we both hate the smell of most sunscreens.
If you have sensitive skin or are looking for a more natural product, I highly recommend this!
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Reviewed in the United States on March 25, 2026
★★★★★ 5
Great reef-safe, easy-to-apply sunscreen (comparison of three ThinkSport products)
Size: 6 Ounce (Pack of 1)
I will start by saying that I am used to traditional non-mineral sunscreens and tend to only buy physical/mineral-based sunscreens when I'm going to be in a tropical ocean and need something reef-safe, though more and more kids sunscreens are shifting to mineral-based. After quite a bit of research I settled on ThinkSport for my family's recent trip to Hawaii, and I'm very pleased with it. I brought and used three types of Thinksport sunscreen: Thinksport Kids lotion (yellow/orange and teal squeeze bottle), Thinksport adult lotion (Black, white, and orange squeeze bottle), and Thinksport adult spray (black, white and orange spray bottle). All protected us from sunburn and rubbed in to be invisible as long as skin was dry and free of visible hair (visible whiteness persisted around my husbands facial stubble and chest hair). The only difference between the kids lotion versus the adult lotion is scent--the kids version has a fruitier, more tropical but not-too-strong scent (possibly coconut and something else? it was hard to identify) that I found unobjectionable (though one if my kids wasn't a fan), and the adult lotion had a lighter, less sweet, more citrus-y scent (my son though maybe lime and mint, but I'm not sure that's accurate) that I LOVE. The adult spray had no discernible scent, so I suspect the kids spray might be fairly unscented as well. The easiest to apply by far was the spray--it sprays on white, but feels very light, rubs in easily, and becomes invisible quickly. I was a little worried about whether it could possibly be effective given how light and thin it felt, but it was! The lotions weren't thick for a lotion (but much thicker than the spray) and went on easily, but took more rubbing in to disappear than the spray (this is consistent with my experience with lotion vs spray in traditional sunscreens as well). Invisibility was harder to get after a second or third application or when skin was damp, but all-in-all I found all three Thinksport products to be superior to other mineral-based sunscreens I have tried in the past. In the future I'll probably buy the adult thinksport spray for my kids and husband (who all much prefer a spray-on sunscreen) and the adult thinksport lotion for myself (because I love the scent and don't mind rubbing it in).
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Reviewed in the United States on April 17, 2023
★★★★★ 5
Dependable sun protection for active days outdoors
Size: 6 Ounce (Pack of 1)
I like how this sunscreen provides reliable broad-spectrum protection and holds up well during outdoor activities. The zinc oxide formula gives me peace of mind, and I appreciate that it's water resistant and reef-safe. It applies smoothly for a mineral sunscreen and stays on effectively during sports, swimming, and long days in the sun. Overall, it's a trustworthy option for anyone looking for strong sun protection.
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Reviewed in the United States on June 8, 2026