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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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4.7 ★★★★★
Based on 20 reviews
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Product Reviews
★★★★★ 5
Good intermediate smart toy
This has become one of my puppy's favorite boy's. He is so smart and flustered through the beginning interactive toys quick. I got him this one next. The design is good. He did get through some ok, but got frustrated early with the harder ones. Started flipping and chewing it. It wasnt easy to chew through but did snap a few hinges. Stubborn dog, but still enjoying it. . It moves easily and does stimulate him to think.
Great toy especially for smart dogs. Mine is just too impatient and growls at it.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on March 18, 2026
★★★★★ 5
Great toy
Color: Yellow
My dog loved this! Kept him entertained for some time. Only problem I had was trying to unscrew the ends after he was done so I could clean it (my dog slobers a lot lol). With him playing and trying to chew it made the ends tighten more. But over all great toy!
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Reviewed in the United States on May 25, 2026
★★★★★ 5
Fun toy!
Color: Yellow
It was funny to see how my cat figured out how to roll the toy to get treats out. Great idea that makes your pet work a little for a reward.
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Reviewed in the United States on June 3, 2026
★★★★★ 4
Cool toy
Color: Yellow
Its pretty cool & drops a piece of kibble about every 5'. It rolls easily on my wood floors & has rubber type ends so it doesn't bang up my wood floors. Only downside is my golden retriever doesn't get how to use it yet. Still working on it.
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Reviewed in the United States on April 28, 2026
★★★★★ 5
BoYoYo Interactive Dog Puzzle Toy – Daily Slow Feeder That Actually Keeps My Dog Engaged
Color: Yellow
This is one of the few dog products I’ve repurchased—and I use it every day as both a slow feeder and enrichment toy.
I have a high-energy dog, and most puzzle toys get solved in seconds. This one is different because of the rolling, unpredictable movement. Instead of just opening compartments, my dog has to actively work to release the food, which keeps him engaged much longer.
What stands out:
• Works as a true slow feeder: I use this for full meals (not just treats), and it significantly slows eating compared to a bowl.
• Higher engagement than standard puzzles: The movement adds a challenge that flat boards don’t have.
• Holds up to regular use: I use this daily for meals, and it’s held up well. My dog hasn’t tried to destroy it, but it’s been more durable than most enrichment toys I’ve used.
Things to know:
• It can be a bit noisy on hard floors because it rolls.
• You need a little open space for it to work properly.
• Not indestructible—best used with supervision if your dog is a heavy chewer.
Bottom line:
If you’re looking for something that actually doubles as a slow feeder and keeps your dog mentally engaged, this is one of the few that’s held up in our daily routine. The fact that I’ve bought it twice says everything.
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Reviewed in the United States on February 24, 2026
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