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Description
Mouse F5 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 V (F5) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by peroxidase (HRP) catalysis and to yellow by acid. The intensity of the color is positively correlated with the amount of Coagulation Factor V (F5) 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 V ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Coagulation factor V (F5) is a protein in the coagulation system composed of six domains: A1-A2-B-A3-C1-C2. Unlike most other coagulation factors, it lacks enzymatic activity and instead functions as a cofactor. Defects can lead to bleeding tendencies, while some mutations (most notably F5 Leiden) predispose to thrombosis. The F5 gene is located on chromosome 1 (1q24). It is genomically associated with the multicopper oxidase family and shares homology with coagulation factor VIII. | |||||||||||||||||||||||||||||||||
| 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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4.4 ★★★★★
Based on 15 reviews
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Product Reviews
★★★★★ 5
This is What You Need on Those Cold Winter Nights!
Color: White, Size: Throw-50"x60"
This Touchat 1000GSM Faux Fur Throw Blanket is absolutely incredible and one of the softest, coziest blankets I’ve ever owned! The thickness and quality are immediately noticeable the moment you touch it. It feels luxuriously plush, warm, and incredibly comforting.
The faux fur texture is beautiful and looks very high-end, adding a stylish and elegant touch to the room while also being amazingly functional for cold nights. The blanket has a nice heavy weight to it that makes it feel extra cozy and relaxing without being too overwhelming.
I love curling up with this blanket while watching TV, reading, or sleeping. It provides outstanding warmth and comfort and feels like wrapping yourself in pure softness. Even after use, it maintains its fluffy texture and doesn’t shed like some faux fur blankets do.
The quality truly exceeded my expectations. The stitching is well done, the material feels durable, and it looks far more expensive than it actually is. It would also make an amazing gift because it feels so luxurious and comforting.
Highly recommend this blanket to anyone wanting something extra warm, thick, soft, and beautiful for their home. Absolutely worth every penny!
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Reviewed in the United States on May 10, 2026
★★★★★ 5
Stunning, soft & well made
Color: White, Size: Throw-50"x60", Color: White, Size: Throw-50"x60"
This blanket is beautiful! Soft on the back side and the feel of the fur is thick and real feeling. Looks absolutely stunning with my velvet pillows also from Amazon. Blanket is a good size for an adult and warm too. The overall look is very high end IMO. I see this blanket as being well made and something that will last. Overall I'm super pleased and would recommend to a friend! I will probably buy another for the bed.
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Reviewed in the United States on December 30, 2025
★★★★★ 5
Luxury-so much luxury
Color: White, Size: Throw-50"x60", Color: White, Size: Throw-50"x60"
Love this blanket. Very warm,thick and heavy,lux fur feels real&soft even after washing(4 washings to date). One side it the fur,the reverse side is a suede-like material that helps it adhere to the bed couch whatever. In between is a thick duvet like filling. The only caveat and it is minor is that the fur is quite slippery. If used as a tire over a couch your guest will be sliding off 😁. Otherwise this throw is a dream. Well worth the money
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Reviewed in the United States on December 29, 2025
★★★★★ 5
Perfect
Color: White, Size: Throw-50"x60"
Warm, soft, big and gorgeous. This is exactly the blanket I was looking for to make my club chair look chic while also being useful, not simply decorative.
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Reviewed in the United States on April 2, 2026
★★★★★ 5
Lushes and warm
Color: White, Size: Throw-50"x60"
This throw is fabulous. Every throw I have
Purchased from Amazon is soft, thick fur,
Perfect color, and lasts. I am tickled pink with the thickness of this throw.
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Reviewed in the United States on February 19, 2026
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