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Description
Mouse F8 ELISA KitProduct Specification Usage Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High precision pipette and gun tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37 constant temperature box 4. Distilled water or deionized water Sample processing and requirements: 1. Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4 overnight, then centrifuge at 1000g for 20
Product Specification
| Usage |
Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High-precision pipette and gun tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37℃ constant temperature box 4. Distilled water or deionized water Sample processing and requirements: 1. Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4℃ overnight, then centrifuge at 1000×g for 20 minutes, and take the supernatant, or store the supernatant at -20℃ or -80℃, but avoid repeated freezing and thawing. 2. Plasma: Collect specimens using EDTA or heparin as an anticoagulant and centrifuge at 1000 × g for 15 minutes at 2-8°C within 30 minutes of collection. The supernatant can be assayed immediately or stored at -20°C or -80°C, but avoid repeated freeze-thaw cycles. Pre-Assay 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 stand for 15 minutes to completely dissolve, then gently mix (concentration 40 ng/mL). Then dilute to the following concentrations: 40 ng/mL, 20 ng/mL, 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, and 0 ng/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each. Pipette 500uL of the 40ng/mL standard working solution into the first EP tube and mix thoroughly to make a 20ng/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 (F8) 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 (F8) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Coagulation Factor VIII ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Factor VII (I 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 clotting disorder. Factor VIII is produced in the sinusoidal cells of the liver and the endothelial cells outside the liver, and is found throughout the body. This protein circulates in the blood in an inactive form, bound to another molecule called von Willebrand factor, until an injury occurs that damages a blood vessel. In response to injury, factor VIII becomes activated and dissociates from von Willebrand factor. The active protein (sometimes written as factor VIIIa) interacts with another clotting factor called factor IX. This interaction triggers a cascade of additional chemical reactions that form a blood clot. | |||||||||||||||||||||||||||||||||
| 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 | 0.62-40 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma |
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4.9 ★★★★★
Based on 27 reviews
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Product Reviews
★★★★★ 1
Squeaker inside toy breaks after 2 days
Size: Mini, Color: Gator (Pink)
Bought for my 8 lb chihuahua for Christmas. I have 3 little girls. The youngest loves her toys and of course she took this one over immediately. She just loved everything about it. The material on the gator is amazingly durable and I believe would hold up for a good while. The squeaker inside the toy however, broke after 2 days of my girl playing with it. It's now a noiseless toy that she does not like too much as it does not squeak because it only had the one squeaker in it.
I would recommend that 2 squeakers be inserted into this gator. I would not purchase again due to the longevity of the squeaker...only lasted 2 days.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on January 5, 2026
★★★★★ 5
Go dog toys are the best!
Size: Large, Color: Dinos Frills (Pink)
These are the only stuffed toys I buy for my Pittie because they are so well made. Even when she unstuffs them, she plays with them forever!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 16, 2026
★★★★★ 5
Amazing for Heavy Chewers!
Flavor Name: REAL Jack Salmon, Size: Puppy (2-Pack)
If you have a dog that destroys every toy in minutes, Benebone is a total game changer. These chews are incredibly durable and actually last—especially for strong or aggressive chewers. They’re made with real flavors like bacon, chicken, and peanut butter, which dogs absolutely love, and you can tell because they keep coming back to it again and again.
What I love most is that they keep my dog busy and satisfied without the mess or smell of traditional bones. They’re also thoughtfully designed with shapes that are easy for dogs to grip and chew comfortably.
That said, like any hard chew, it’s important to supervise and replace it if it gets worn down—but overall, this is one of the best long-lasting chew toys out there.
Highly recommend for anyone with a serious chewer—worth every penny! 💛🐶💛
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Reviewed in the United States on March 20, 2026
★★★★★ 5
Excellent quality chews. Tiny Puppy size is perfect for adult toy breed dogs.
Flavor Name: REAL Bacon, Size: Puppy (2-Pack)
I bought the Tiny Puppy Bacon flavor 2-pack package of these, and my dog and his sister are obsessed with them. They chew on them multiple times every day. They seem to be fairly durable, although I expect they will break down over time, which is okay as it’s a chew. I like that they stay intact, with no chunks breaking off for the dogs to swallow. The dogs do ingest a small amount as the material breaks down, but I always supervise them when they have these, and there haven’t been any problems. It’s difficult to find good chews for toy breed dogs like our 12-pound Shih Tzus, so I was happy to find these. They are pricey, but I believe they are worth it. The dogs love to carry them around, and my little guy even plays with his chew sometimes like it’s a toy.
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Reviewed in the United States on February 20, 2026
★★★★★ 5
Durable, engaging, and dog approved
Flavor Name: REAL Bacon/REAL Maplewood, Size: Puppy Tiny (2-Pack)
These chew toys are great for keeping dogs entertained and engaged, especially for gentle chewers. The quality feels strong and they hold up well compared to many other toys. The real bacon scent keeps dogs interested and coming back to them often. I’ve ordered multiple packs over time for my own dogs and even gifted them to others because they’re so well liked.
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Reviewed in the United States on May 2, 2026
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