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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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4.6 ★★★★★
Based on 15 reviews
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Product Reviews
★★★★★ 5
Great play sneaker for outside
Size: 1 Little Kid, Color: Fuchsia
These sneakers fit my daughter’s feet perfectly and she loves the color. I trust the Saucony brand as I’ve owned several pairs. They are well made and this pair is an excellent option for play sneakers. My kids are outside a lot and I need a sneaker that can handle running, climbing and occasionally getting wet. There is generous breathable mesh and the shoes dry quickly.
My daughter has slightly wider feet and these fit comfortably. I also appreciate that they are Velcro so can be put on or taken off quickly because we all know kids are impatient when it comes to playtime.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 19, 2026
★★★★★ 5
Great everyday sneaker
Size: 1 Little Kid, Color: Fuchsia
I bought these for my kid and honestly didn’t expect to like them as much as I do, but they’ve been great so far. They’re lightweight, easy to get on, look cute, fit well, and actually seem comfortable enough that there’s been zero complaining (which is saying a lot).
The grip on the bottom is solid, and they’ve held up really well with everyday wear, running around, playground, all of it. I also like that they’re breathable, so no sweaty feet at the end of the day.
They’re not the most “fancy” sneaker out there, but for the price, the quality feels really fair. If anything, I wish they came in even more color options because we’d probably buy another pair.
Overall, really good everyday shoe. If you just need something reliable, comfortable, and kid-proof, these definitely do the job.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 21, 2026
★★★★★ 5
You've Found Your Daughter's Match! Superior Sneakers for Girls!
Size: 1 Little Kid, Color: Fuchsia, Size: 1 Little Kid, Color: Fuchsia
Super-cute, stylish sneakers meet cushiony comfort at a very reasonable price. These sneakers have it all!
The bed and sole of these sneakers provide significant shock-absorption making them ideal for serious athletics or a long day out at the theme park. Our daughter went from a Keds-type sneaker to these and she was beaming 😁!
In addition to their comfort and ppearance is the fact that they fasten and unfasten with ease. Any child can put them on and take them off. The laces stretch and the Velcro is highly adjustable. Everything has been constructed with integrity.
They are, also, washable so you can continue to maintain their crisp, vibrant colors and keep them looking new.
They fit true-to-size and can accodimate somewhat wide feet as well.
There are only positive things to say about these 5+ stars!
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Reviewed in the United States on April 11, 2026
★★★★★ 5
Nice to have a wider shoe! Nice color and seems to be good quality so far!
The fit ran a little bit small compared to Nike brand. Size chart at a shoe store I felt he needed a 1.5 (with room to grow still), ended up ordering a 2.5 which may have been a tiny big but works for his feet to grow. Love that it's wider. Cute color
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Reviewed in the United States on January 18, 2026
★★★★★ 5
Cute, durable kids' shoe at a great value
Size: 11.5 Big Kid, Color: Navy/Citron
These worked out great for our 5 year old with flat feet - he absolutely loves them, says they are very comfortable and help him "run FAST!" The colors look super cute, I'm very pleased with the durability over several months (particularly, the hook & loop closure has held up better than other brands we've tried...), and they seem to offer great support while not being too heavy. Highly recommend, and will buy again.
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Reviewed in the United States on August 26, 2025
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