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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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4.5 ★★★★★
Based on 27 reviews
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Product Reviews
★★★★★ 4
Four Stars: Insightful and Impactful Reading
Format: Hardcover
This book provides clarity in human relationships pertaining to workplace, family, and social circles. Author Mel Robbins explains the human tendency of needing to belong and bond, having personal freedoms and space to make informed choices, and providing coping strength to not feel over burdened due to work or perceived social obligations or human stereotypes. "Let Them", "Let Me" is a series of strategies/skills that help one regain focus on finite time and energies to have more productive personal relationships and work outcomes. The book teaches one to de-prioritize things and channel emotions into actionable goals to set healthy boundaries in selection of friends, career paths, and effectively resolving conflicts. Some chapters are overly redundant (and precludes a fifth star rating); however, it's a worthwhile book that definitively helps me set more impactful goals in 2026 and beyond.
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Reviewed in the United States on January 31, 2026
★★★★★ 5
HIGHLY RECOMMEND
Format: Audiobook
I love Mel Robbin’s and this audiobook gave me a different perspective on my life. I would highly recommend it’s so easy to understand and it’s so enjoyable and enlightening to listen to.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 15, 2026
★★★★★ 5
Book of life
Format: Hardcover
This book is a fantastic read, I think this is a book everyone should read. Very insightful and informative book for life.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 15, 2026
★★★★★ 5
NON FICTION IS A GREAT WAY TO LISTEN WHILE YOU DRIVE! TRY AUDIBLE!
Format: Audiobook
LET THEM
AUDIBLE NON-FICTION INSPIRATIONAL BOOK ON LISTENING TO MEL ROBBINS MOTIVATIONAL ADVICE MAKES MY DRIVING DESTINATIONS ENJOYABLE! I had something like 24 AUDIBLE CREDITS that have built up because I'm a visual person that loves to read. I've had the membership for over ten years and I only like certain NON FICTION SELF DEVELOPMENT books which I listen to Memoirs, Self help, but never fiction books, because my imagination is more vivid when I read. I always either listen to my podcasts on Spotify or NON-FICTION on AUDIBLE which is owned by Amazon. I can't drive anywhere without my Podcasts or very few Non-Fiction Titles that interest me, but I have had those credits and I love MEL ROBBINS so I decided to try it on AUDIBLE. My friends who read that listen to fiction on AUDIO understood why I bought the same fictional title and I had already loved reading it on my Kindle. It was a favorite lifetime reading experience and I own many of the same KINDLE & AUDIBLE versions but I noticed that fiction book that it was one of the most beautifully written books that when I went to listen to it, I quickly gave up because it was bland. I have been told by more frequent readers like myself who listen to the Audio version and recommend it to me on Audio since they enjoyed it better have said listening to fiction is a skill that you have to develop, so I stick to NON FICTION and I need to listen to something while I'm driving so I buy the Kindle Version & the AUDIBLE and it makes my driving to destinations shorter.v That's why I have Spotify because there's two favorite podcasts but I pay extra for no commercials and now my top favorite podcast has commercials which I 'm considering cancelling because the reason I pay extra is because the commercials drive me crazy so now Spotify says it's up to the podcast. The reason I've been a member since 2018, is because it is synced to the Bluetooth, in my car and the podcasts don't require Cell towers so you don't get the buffering that I do with Audible in areas where there aren't Cell towers which we call them dead zones. I'm recommending this AUDIBLE PURCHASE called, "LET THEM," because MEL ROBBINS is a new motivational author to me, and she wrote the book which she has some fascinating inspirational content, that I have learned some ideas to improve my life. She is a fantastic narrator in her soothing voice. One nice thing about AUDIBLE is they are extremely friendly and if you don't like the AUDIO BOOK Listening experience, you can return it, which you can't do with the KINDLE Books. so I have had to return something only twice in fifteen years, since I usually love what I purchase. I do recommend this book on AUDIBLE to listen to or if you don't have a membership they used to have a free trial and a free book if you're a new member. But I'm not sure, but however medium you choose this is a FAVORITE and it's a bestseller that I can promise you will love it, as well. It has a very high rating of tens of thousands of five star ratings which is even more evident that it's a Spectacular choice!
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Reviewed in the United States on July 24, 2025
★★★★★ 5
Let Them and Let Me, is what I needed!
Format: Hardcover
I wish that I could give this book much more than 5 stars. I absolutely loved reading it. The book is so interesting and the information is useful and makes a lot of sense. I have been reading the book since I received it on March 5, 2026 and it was very hard for me to put it down. I am now on the concluding chapter "Your Let Me Era Is Here." I feel a little sad that I am finishing the book because I want to know more. However, I decided to practice the suggestions and theories the author stated in the book to see if they really worked. The 5-4-3-2-1 suggestion was used anytime there was something that I didn't feel motivated to do, and it worked. I felt better afterwards. The "Let Them" and "Let Me" theories have worked also. But, I realized it is going to take time for me to make the needed changes consistently. Still, I am very happy that I purchased and read this book. It is definitely worth the money and time spent reading and making application of the information. It boosted my confidence!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on March 29, 2026
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