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Description
Biotin 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: The detection range of the kit is not equivalent to the concentration range of the analyte in the sample. Before the experiment, it is recommended to estimate the analyte concentration in the
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: The detection range of the kit is not equivalent to the concentration range of the analyte in the sample. Before the experiment, it is recommended to estimate the analyte concentration in the sample based on relevant literature and conduct preliminary experiments to determine the actual concentration in the sample. If the analyte concentration in the sample is too high or too low, dilute or concentrate the sample appropriately. If the sample type is not listed in the instructions, it is recommended to conduct a preliminary experiment to verify the validity of the test. Serum: Collect whole blood in a serum separator tube and place it at room temperature for 2 hours or at 2-8°C overnight. Then centrifuge at 1000×g for 20 minutes and remove the supernatant. Alternatively, store the supernatant at -20°C or -80°C, but avoid repeated freezing and thawing. Plasma: Collect the specimen using EDTA or heparin as an anticoagulant and centrifuge at 1000×g for 15 minutes at 2-8°C within 30 minutes of collection. Remove the supernatant and analyze. Alternatively, store the supernatant at -20°C or -80°C, but avoid repeated freezing and thawing. Tissue homogenate: Rinse the tissue with pre-chilled PBS (0.01M, pH 7.4) to remove residual blood (lysed red blood cells in the homogenate may affect the measurement results). Weigh and mince the tissue. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1g of tissue sample to 9mL of PBS. The specific volume can be adjusted according to experimental needs and recorded. It is recommended to add protease inhibitors to the PBS) in a glass homogenizer and grind thoroughly on ice. To further lyse the tissue cells, the homogenate can be sonicated or repeatedly freeze-thawed. Finally, centrifuge the homogenate at 5000×g for 5-10 minutes, and collect the supernatant for analysis. Cell Lysis Buffer: Adherent cells are gently washed with ice-cold PBS, then trypsinized and harvested by centrifugation at 1000×g for 5 minutes. Suspension cells can be harvested directly by centrifugation. The harvested cells are washed three times with ice-cold PBS and resuspended in 150-200µL of PBS per 1×10^6 cells (it is recommended to add protease inhibitors to the PBS; if the cell count is very low, the PBS volume can be reduced appropriately). Disrupt the cells by repeated freeze-thaw cycles or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and remove the supernatant for testing. Cell culture supernatant: Centrifuge at 1000×g for 20 minutes, remove the supernatant, and test immediately. Alternatively, store the supernatant at -20°C or -80°C, but avoid repeated freezing and thawing. Other biological samples: Centrifuge at 1000×g for 20 minutes, remove the supernatant, and test immediately. Sample Appearance: The sample should be clear and transparent, and suspended matter should be removed by centrifugation. Sample Storage: Samples collected for testing within one week can be stored at 4°C. If testing is not possible, aliquot the sample into single-use aliquots and freeze at -20°C (for testing within one month) or -80°C (for testing within six months). Avoid repeated freezing and thawing. Hemolysis of the sample can affect the final test results, so hemolyzed samples are not suitable for this test. Preparation for the test: 1. Remove the reagent 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, 5 ng/mL, 1.25 ng/mL, 0.313 ng/mL, 0.078 ng/mL, 0.02 ng/mL, 0.005 ng/mL, and 0 ng/mL. Serial dilution method: Take seven EP tubes and add 600 μL of universal diluent to each tube. Pipette 200 μL of the 20 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 5 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 HRP-SA working solution: 15 minutes before use, centrifuge the concentrated HRP-SA at 1000×g for 1 minute. Dilute 100× concentrated HRP-SA to a working concentration of 1× with universal diluent (e.g., 10 μL concentrate + 990 μL universal diluent). Prepare and use immediately. 4. Prepare 1× Wash Buffer: Dissolve 10 mL of 20× Wash Buffer in 190 mL of distilled water. (Concentrated Wash Buffer removed from the refrigerator may crystallize, which 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 them to 4°C. 2. Sample Addition: Add 50 μL of sample or standard of varying concentrations to the corresponding wells. Add 50 μL of Universal Diluent to the blank wells, followed by 50 μL of HRP-SA working solution to each well. Cover with film and incubate at 37°C for 60 minutes. (Recommendation: Dilute the sample to be tested at least 1x with universal diluent before adding it to the ELISA plate for testing. This will reduce the impact of matrix effects on the test results. The final calculation of the sample concentration should be multiplied by the corresponding dilution factor. It is recommended to set up duplicate wells for all samples to be tested and standards.) 3. Washing: Discard the liquid and add 300 μL of 1x wash buffer to each well. Let it stand for 1 minute. Shake off the wash buffer and pat dry on absorbent paper. Repeat this process 5 times (a plate washer can also be used for washing). 4. Adding substrate: Add 90 μL of substrate (TMB) to each well, cover with sealing film, and incubate at 37°C in the dark for 15 minutes. 5. 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: Interpreting Results: 1. Calculate the average OD value of the standard and sample replicates and subtract the blank OD value as a correction value. Plot the standard curve for 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 when calculating the sample concentration. |
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| Sensitivity | 0.001 ng/mL | ||||||||||||||||||||||||||||||
| Theory | This kit uses a competitive enzyme-linked immunosorbent assay (ELISA). Samples, standards, and HRP-labeled streptavidin (HRP-SA) are sequentially added to microwells pre-coated with biotin antibodies. 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 color intensity is negatively correlated with the biotin content in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | ||||||||||||||||||||||||||||||
| Source | All | ||||||||||||||||||||||||||||||
| Synonym | General Biotin ELISA Kit | ||||||||||||||||||||||||||||||
| Detection Type | Competition Law | ||||||||||||||||||||||||||||||
| Composition |
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| Background | Biotin (vitamin B7) is a vitamin found in foods such as eggs, milk, and bananas. It is an essential component of enzymes in the body that break down fats, carbohydrates, and other substances. Biotin has been used in alternative medicine as an effective adjunct to treating or preventing biotin deficiency. Biotin deficiency can be caused by malnutrition, rapid weight loss, long-term tube feeding, and other medical conditions. Biotin is a water-soluble B vitamin. Consuming large amounts of biotin as a dietary supplement results in absorption and subsequent excretion in the urine as biotin. Consuming biotin as part of a normal diet results in urinary excretion of biotin and biotin metabolites. | ||||||||||||||||||||||||||||||
| 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 | 0.005-20 ng/mL | ||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.6 ★★★★★
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Product Reviews
★★★★★ 5
Perfect replacement wipers
Size: 26"+14"+10-1(PTB)
These are fantastic and fit my car perfectly. I was a little worried but they went on easily and were the perfect replacement. They fit against the glass and don't cause any smearing. I was thrilled to get them so quickly because we had another storm coming and these worked fantastically. I can see out of my windshield. If you're on the fence about these, go ahead and order, they work great.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on March 18, 2026
★★★★★ 5
Great replacement wiper blades.
Size: 26"+17"+12JP(HOOK)
These were great wiper replacement for my car, very easy to install and way cheaper that buying them at a auto parts store.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 27, 2026
★★★★★ 5
Impressive performance at an impressive price
Size: 26"+17"+14BP(HOOK)
I had one damaged wiper, so I decided to replace all 3 on my Subaru Outback. I have read a lot of positive information regarding "silicone" wipers, so I settled on this complete kit as it was a good value for the price and received many positive reviews.
They come well packaged, yet I was concerned that the longest blade had to be bent significantly to fit in the odd shaped box. However, once installed, that blade straightened out and conformed snuggly to the contour of the windshield.
True to the claim, it does take about 30 seconds to install each blade (that is if you understand the diagram and attempt to do so correctly). I had no issues and each blade snapped securely in place. They appear to be well made and durable. The design is clean and they look good on my vehicle.
I've been using them for 2 months now and they work well. They clear the windshield without streaking or chatter. They seem to work as well as other types I have used. The ultimate test will be how long they last. So far, I am very pleased with my choice.
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Reviewed in the United States on April 21, 2026
★★★★★ 4
Easy Installation Wipers!
Size: 26"+19"+14BP(HOOK)
Just installed them on my Outback, simple straightforward install. The instructions were simple and they seem to fit well. These wipers were more compatible than the old ones, and the appearance look like they are original!
When I purchased these wipers I thought I had purchased the Rain-X wiper replacements, but they were not. These wipers seem to work when I used the windshield cleaner with the wipers. The windshield cleaning seems effortless and without any noise.
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Reviewed in the United States on May 21, 2026
★★★★★ 5
Great Wiper Blades, Great Price
Size: 26"+18"+16BP(D-PTB)
Perfect fit for my 2020 Subaru Outback. The blades were very well packaged and arrived in perfect condition. They clean perfectly. I have only been using them for a month, so I can’t speak to longevity. They cost less than one third the premium blades cost at the auto parts store, and work just as well.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 8, 2026
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