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Description
Human TNFRSF1B 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 the specimen using EDTA or heparin as an anticoagulant. Centrifuge the specimen at 1000 × g for 15 minutes at 2-8°C within 30 minutes of collection. The supernatant can be assayed or stored at -20°C or -80°C, but avoid repeated freezing and thawing. 3. 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 the tissue and mince it. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1 g of tissue sample to 9 mL 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 sonicated or repeatedly frozen and thawed. Finally, centrifuge the homogenate at 5000 × g for 5-10 minutes, and the supernatant can be assayed. 4. 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 10 ng/mL). Then dilute to the following concentrations: 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 ng/mL, 0.15625 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 10ng/mL standard working solution into the first EP tube and mix thoroughly to make a 5ng/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 capture antibody against Tumor Necrosis Factor Receptor Superfamily, Member 1B (TNFRSF1B). 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 Tumor Necrosis Factor Receptor Superfamily, Member 1B (TNFRSF1B) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Tumor Necrosis Factor Receptor Superfamily, Member 1B ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Tumor necrosis factor receptor superfamily member 1B (TNFRSF1B), also known as tumor necrosis factor receptor 2 (TNFR2) and CD120b, is one of two membrane receptors that bind tumor necrosis factor-α (TNFα). Like its counterpart, tumor necrosis factor receptor 1 (TNFR1), the extracellular region of TNFR2 consists of four cysteine-rich domains that bind to TNFα. TNFR1 and TNFR2 have distinct functions when bound to TNFα due to differences in their intracellular structures, such as the lack of a death domain (DD) in TNFR2. It has been shown to interact with TRAF2 and TTRAP. | |||||||||||||||||||||||||||||||||
| 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 | 0.15-10 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenates and other biological fluids |
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4.4 ★★★★★
Based on 85 reviews
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Product Reviews
★★★★★ 5
The BEST water bottle on the market!
Color: Denim, Size: 32 Ounces
I am in love with this water bottle! I take it everywhere with me, and I love that it has a built-in straw. The color is perfect as well. It motivates me to get my water intake in, and it helps me meet my fitness goals. It is easily washed in the dishwasher, and I don't have to worry about it breaking. It keeps the temperature of my water perfect- whether I add ice or not. It is sturdy but not too heavy.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 26, 2026
★★★★★ 5
Excellent Summer Resource for Kids!
Format: Paperback
I purchased the "Summer Bridge Activities 2nd to 3rd Grade Workbook" for my 8-year-old, and it has exceeded my expectations in every way. This comprehensive workbook is a fantastic tool to keep children engaged and learning during the summer break. My sibling is an elementary school teacher and introduced me to these books when my son was in Kindergarten and I honestly have not looked back. My LO loved a good challenge so we typically do multiple days to ensure we finish it in its entirety during the summer. I'm constantly recommending these books every summer as I have not yet found anything to replace it.
Content Quality:
The workbook covers all essential subjects: math, reading comprehension, writing, science, social studies, and even fitness. Each section is well-organized and filled with age-appropriate activities that challenge and entertain. The variety of exercises ensures that my child doesn't get bored, and the progressive difficulty helps build confidence.
Educational Value:
The math and reading comprehension sections are particularly strong, with clear explanations and plenty of practice problems. The writing prompts encourage creativity and critical thinking, while the science and social studies sections introduce interesting facts and concepts. I appreciate the inclusion of fitness activities, promoting a healthy lifestyle alongside academic learning.
Flash Cards:
The included flash cards are a wonderful bonus. They are durable, colorful, and perfect for quick review sessions. My child enjoys using them, and they make reinforcing concepts fun and interactive.
Ease of Use:
The workbook is user-friendly with clear instructions for both parents and kids. It's designed to be self-paced, which means my child can work independently or with minimal guidance. The answer key at the back is helpful for quick checks.
Engagement:
What sets this workbook apart is its ability to make learning enjoyable. The activities are engaging, and the themes are relevant and interesting. My child looks forward to workbook time each day, which is a huge win for any parent trying to prevent the summer slide.
Overall, the "Summer Bridge Activities 2nd to 3rd Grade Workbook" is an excellent investment in your child's education. It's thorough, fun, and well-designed to keep young minds sharp over the summer. I highly recommend it to any parent looking to provide a balanced and enriching educational experience during the school break.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on June 13, 2024
★★★★★ 5
Good quality for price
Format: Paperback
This was a good book for the summer. It has a variety of activities so my child didn't get bored. It also has some fun "outside" only ideas to get them up and moving. It was a good bridge book that she could (mostly) do independently. However, as they progress through it, parents should be there to help and reiterate different concepts. It was well worth the price and helped the transition go smoothly.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on November 12, 2025
★★★★★ 5
Perfect To Keep A Youngster's Mind Active Over The Summer
Format: Paperback
Simply put, this is a great activity book to help your child stay mentally active when out of school for the summer. It is not intended to be a full school curriculum, but instead a way to keep children mentally active during the summer break. We have a child leaving 2nd and heading to 3rd grade and were concerned about her skills being lost over the summer. While we do work with her to increase her skillset, we were more concerned about the atrophy that happens when not in school than jumping her ahead. It can be a bit tough for kids returning to school if they go 2-3 months with little book work, so we wanted to help keep her fresh but not fill her summer with full days of 'school' work.
This book is a solid mix of skills with some being easy and others more of stretch. The easy and stretch items will vary with the schooling your child gets and their abilities. I suspect most kids heading to third grade will find most of the work in this book at least a strong refresher and potentially help move them in some areas. This is setup to be a roughly 15 minute daily regime but we have enhanced it with some other work. We have math and science specific activity books as well as a reading workbook that we rotate into the mix. We also keep track of words she has a harder time spelling and use them make a 10 spelling word weekly review and test (review for 4 days and test on the fifth). Most of these topics are in the bridge activities book, but we felt a little extra attention would not hurt and some variety in what she is working on keeps things fresh. We do spelling, reading/writing and a couple 'pages' of work each weekday which comes out to about a 45-60 minute batch of work. The Summer Bridge Activities books is a constant in the mix with a page used almost every day and the other work rotated.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on July 5, 2017
★★★★★ 4
Great summer workbook!
Format: Paperback
So far an easy book compared to the Argo Summer Academy one that was too challenging. I don't like the way the pages are perforated they fall out of the book too easily. I feel like it is a good balance of keeping up skills already learned and softly introducing a little bit of challenge. I like how each day has math and reading. Easy to keep skills up with only 15 minutes of work a day.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on June 22, 2025
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