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Description
Rat TFR2 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 preparation and requirements: Tissue homogenization: Rinse the tissue with pre chilled PBS (0. 01M, pH 7. 4) to remove residual blood (lysed red blood cells in the homogenate will affect the measurement results). Weigh and
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 preparation and requirements: Tissue homogenization: Rinse the tissue with pre-chilled PBS (0.01M, pH 7.4) to remove residual blood (lysed red blood cells in the homogenate will 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 tissue cells, the homogenate can be ultrasonically disrupted or repeatedly frozen and thawed. Finally, the homogenate is centrifuged at 5000×g for 5-10 minutes and the supernatant is collected for analysis. Other biological fluids: Centrifuge at 1000xg for 20 minutes, remove the supernatant, and test. Pre-test 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 it stand for 15 minutes to completely dissolve, then gently mix (concentration is 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 7 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 directly as a blank well; there is no need to aspirate the liquid from the penultimate tube. See the figure below for details. 3. Preparation of Biotinylated Antibody Working Solution: 15 minutes before use, centrifuge the concentrated biotinylated antibody at 1000×g for 1 minute. Dilute the 100× concentrated biotinylated antibody to a 1× working concentration using universal diluent (e.g., 10µL concentrate + 990µL universal diluent). Prepare immediately before use. 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 utilizes 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 transferrin receptor 2 (TFR2) 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 transferrin receptor 2 (TFR2) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Transferrin Receptor 2 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Transferrin receptor 2 (TFR2) is a protein encoded by the TFR2 gene. This protein is involved in the uptake of transferrin-bound iron into cells via endocytosis, although its role is less pronounced than that of transferrin receptor 1. This gene, a member of the transferrin receptor family, encodes a single-pass, type II membrane protein with a protease-associated (PA) domain, an M28 peptidase domain, and a transferrin receptor-like dimerization domain. This protein mediates the cellular uptake of transferrin-bound iron, and mutations in this gene are associated with hereditary hemochromatosis type III. | |||||||||||||||||||||||||||||||||
| 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 | Tissue homogenates and other biological fluids |
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4.8 ★★★★★
Based on 28 reviews
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Product Reviews
★★★★★ 5
Great, funny intro to AI
Format: Hardcover
This is a wonderful, humorous introduction to AI that is a fast read packed full of examples. It makes a great gift for friends or family who don’t know much about the field, and I imagine it would be especially interesting to teens. I loved the way she used running jokes to make connections between themes. Shane is clearly on a mission to make AI more accessible.
It’s funny, someone else said they didn’t like this book because it wasn’t enough like Shane’s tear-inducingly-hilarious blog. But here’s the thing: I can read the blog for free! I was actually nervous that I was going to be getting a repeat of the blog in book form, but it was super different. If you’re a computer science scholar, maybe skip this one, but I don’t think that was the audience Shane was trying to reach.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on November 19, 2019
★★★★★ 3
Humorous intro-level book to AI
Format: Hardcover
If you are new to the field of AI, this is a fun read. The author does a fine job breaking down complex topics into bite-size, entertaining sections. However, if you are not new to the field of AI, you'll probably find this book boring and will find more joy in visiting the author's popular website.
Personally, I find this book forgettable and not living up to the hype.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 1, 2020
★★★★★ 5
Captivating storytelling and educational material
Format: Paperback
My ten years old boy loved WW2 in Europe: Great Battles for Boys. He likes to read about war and it his third book from Joe Giorello. It is an engaging and easy-to-understand account of significant World War II moments designed for young boys. It has pictures and maps so they can have a better understanding about the War and through captivating storytelling and educational material, it provides a valuable introduction to this crucial era, promoting a greater comprehension and respect for the experiences and achievements of individuals during that time.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on March 17, 2024
★★★★★ 5
Why are these books so hard to find in libraries and bookstores??!
Format: Paperback
Bought nearly the whole book series after reading the reviews. My 8 and 10 year old boys love these books. My 10 year old read the first book on his way to and from soccer practice. He couldn't put it down and kept telling me what he read. My boys have read all the "Magic Treehouse", "I Survived" and some "I Escaped" books, as well as "Big Nate", "Diary of a Wimpy Kid", etc. BUT these battle books are the best. They really teach about the history of the wars which have shaped our modern times. WHY are these books so hard to find in libraries and bookstores... at least in Maryland? Maybe because "Great Battles for Boys" reeks of toxic masculinity? I really wonder. In any case, great books especially if you want your boys to read serious, informative books that are interesting and entertaining.
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Reviewed in the United States on February 1, 2024
★★★★★ 5
Great for boys AND MOMS
Format: Paperback
Saw an ad for the series online and wanted to buy just one book to try. Received with 2 day Prime. Book was new. So, not only does this book have an incredible, intriguing story that sucked in my 10 year old who hates reading and struggles to read, it had me at Hello. The beginning of the book is filled with recipes! Was totally not expecting that! So for this Momma, under promise-over deliver was the theme here. Purchasing the rest of the books! Also look online for worksheets that parents have put together to go along with these books. Totally worth it. Happy Reading! AND Cooking!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on January 25, 2025
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