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Description
Mouse TNFRSF1A 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: 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: 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. 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. 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. Cell Lysis Buffer: Gently wash adherent cells with ice-cold PBS, then trypsinize and collect cells by centrifugation at 1000 × g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash the collected cells three times with ice-cold PBS and resuspend them in 150-200 μL of PBS per 1 × 10^6 cells (it is recommended to add protease inhibitors to the PBS; if the cell count is very low, reduce the PBS volume 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 collect the supernatant 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. Preparation of standard gradient working solution: Add 1mL of universal diluent to the lyophilized standard, let it stand for 15 minutes to completely dissolve, then gently mix (concentration is 5000pg/mL). Then dilute to the following concentrations: 5000pg/mL, 2500pg/mL, 1250pg/mL, 625pg/mL, 312.5pg/mL, 156.25pg/mL, 78.125pg/mL, and 0pg/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each. Pipette 500uL of the 5000pg/mL standard working solution into the first EP tube and mix thoroughly to make a 2500pg/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 enzyme conjugate are sequentially added to microwells pre-coated with a capture antibody against Tumor Necrosis Factor Receptor Superfamily, Member 1A (TNFRSF1A). 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 1A (TNFRSF1A) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Tumor Necrosis Factor Receptor Superfamily, Member 1A ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Tumor necrosis factor receptor superfamily member 1A (TNFRSF1A), also known as tumor necrosis factor receptor 1 (TNFR1) and CD120a, is a ubiquitous membrane receptor that binds tumor necrosis factor-α (TNFα). Germline mutations in the extracellular domain of this receptor have been found to be associated with a condition called tumor necrosis factor-associated periodic syndrome (TRAPS). | |||||||||||||||||||||||||||||||||
| 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 | 78.12-5000pg/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenates, cell lysates and other biological fluids |
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4.5 ★★★★★
Based on 786 reviews
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Product Reviews
★★★★★ 5
Solid, powerful, rechargeable
Color: Black
Very impressed with its weight, speed and performance. The dial is great
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Reviewed in the United States on May 29, 2026
★★★★★ 4
Very nearly the perfect frother for all your basic frothing & mixing needs
Color: Black
There are a lot of middle-of-the-road frothers out there. I've been through a few of them in my recent search for something that could mix and froth well, without taking up any more outlets in my basement kitchen. Of the three Maestri frothers I've tried so far, this one wins the race by a nose.
Most recently, these Maestri frothers come in basically three versions: A single-speed @ 8000 RPM, a two-speed @8000/5500 RPM, and this stepless variable-speed version. Aside from that, the only real difference in recent version pack-outs is which attachments they come with.
Look over the reviews of the single-speed version and you'll find that while it can and does froth well, it starts at a single, high speed and gets there fast. This makes it pretty easy to spin liquid right out of most common cups and mugs.
There is a two-speed version, but it's harder to find, only comes in one color (Grape Purple), and while it's much better than the Maestri single-speed, it still has a couple of quirks that make this variable-speed version win out.
This mixes and froths whole milk, half-and-half, or heavy whipping cream, or just about anything else very well. Like all frothers, it takes a little time to learn its nuances and nail down the technique, this will definitely get you there.
The best feature of this is easily the speed control. Turn the knob to turn it on at a low speed that's great to get things started, then turn the knob to crank up the speed just enough to do what you need, whether that's mixing or frothing. The low starting speed makes it easy to keep things under control without undue spilling, and the max speed is more than enough to make quick work of getting your froth on.
There are really only two complaints I have with this stepless, variable speed version:
- I'd really like to have a Press On / Release Off button in addition to the Speed Control knob. More than one time have I gone to turn this off, only to spin the knob the wrong way and crank the speed up to ludicrous, sloshing liquid on the counter. Being able to turn it Off just by letting go of the button would be quick and easy. This configuration would allow using a preferred speed right from the start, while still allowing speed to be adjusted on-the-fly when needed.
- Give it a bigger battery. It would cost mere pennies to give this a 2000mAH+ instead of a 1200mAH battery, and I can't think of any reasonable downside to that.
- Give the motor a little more torque. It's fairly easy for the current motor, at any speed, to get bogged down in a thick protein powder mix, or when pressing the frother or other attachment a bit too hard into the bottom or side of the frothing container. A bit more "oomph" would prevent that.
I really like the overall design and features of thes Maestri frothers better than many other, cheaper versions. This variable-speed version is pretty great as it is and probably the one I would recommend over the single- or two-speed, for most people. But I often find myself using two hands -- one to hold it steady, and the other to turn it on and tweak the knob to the desired speed(s) -- for a device that should arguably need only one hand to use. Just a couple of minor tweaks as noted above would make this the overall best frother of its type that I've used.
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Reviewed in the United States on May 23, 2025
★★★★★ 5
Hands down, the BEST handheld frother I've ever used!
Color: Black
Hands down, the BEST handheld frother I've ever used! I got the black one, but any color will have the same results.
Over the past 12 years or so, I've used so many different handheld frothers and they've ALL fallen short of expectations for quality, longevity, power, features, and usability.
I've used everything from cheap, $15 models to $50+ models and everything in-between. Some with fancy attachments for various types of mixing, frothing, beating, whipping, etc. Some with rechargeable batteries, some without. Some AC powered, some with fixed whisks, some with detachable/replaceable whisks, some with stands, etc. NONE have been spectacular. Most broke within 6 months and got tossed. ALL were major disappointments in the end, including the "revered" Zulay models of which I tried several.
I finally found this Maestri House branded, variable speed frother with detachable / replaceable whisks and got one to try. I was literally on Cloud Nine the first time I turned it on. Like, WOW! Not only was it FAST on the highest speed, but it was powerful enough to churn right through milk, eggs, cream, etc, without bogging down like many others.
This thing makes milk froth like a milkshake and Matcha Lattes like nothing else I've ever experienced. I used to have to sift my Matcha, then whisk it in hot water, then froth milk and then blend them together to make the perfect latte. and if the milk was cold (my preference), pre-whisking in hot water was a must to avoid lumps. However, with THIS frother? I literally pour my milk into a tall tumbler, drop in un-sifted Matcha powder, and spin up the Maestri House frother at first on a medium low speed to get it mixed, then jump straight to the highest speed to really whip that milk and match up. After about 30-45 seconds, I've got the thickest, richest, smoothest, most aerated latte around. And NO LUMPS at all!!! What a time and dish saver!
And cleaning? I just run it under hot running water to get the shaft cleaned and then spin it up in a dish with hot running water for a few moments, then spin it on high for a few seconds in the air to dry it off instantly.
Now, I'll be honest here, too. The specs say it can go months on a charge, using it for a couple minutes a couple times daily. Well, I guess it could do that and still spin. But I use it FULL SPEED, churning hard for at least a minute a couple times per day. After about 2 weeks, I can start to notice a speed reduction so I just plug it back in on the charger. All things considered, this is still better than all the other handheld frothers I've used over the years.
After using the heck out of this thing for the past 7 months, I've even decided to start selling these in our Japanese gift shop. The manufacturer is has been very responsive both in customer support (I called them about an issue I thought I was having, and they called me back with a couple hours even though I didn't leave voicemail when they didn't answer right away) and in reseller support. I have to give these guys an A+ for responsiveness and quick resolution when problems might arise.
HIGHLY recommended!!
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Reviewed in the United States on October 17, 2025
★★★★★ 5
Great Addition!
Color: Black
Why did you pick this product vs others?:
I just started a powdered drink routine recently and was having a difficult time mixing it with water. I would consistently end up with small nuggets of mix making it unenjoyable to drink. I knew I needed a mixer and was glad I found this device when I did. It works perfectly! I like the fact that it comes with its own stand and included two mixing heads. It is solid, well-designed, and you can feel the quality when you pick it up. The variable speed of the mixer is perfect for ensuring that everything is blended well. I haven’t used it to “froth” anything yet, at least not on purpose, but I accidentally found out that it can do that well. This is a great addition to our kitchen and is very easy to wash after use. So far…no issues. Highly recommend for those of you looking for the right tool to make sure those protein mixes aren’t full of unmixed nuggets.
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Reviewed in the United States on June 21, 2025
★★★★★ 4
Low foam
Color: Black
Love this frother. It is great quality and the turn speed is amazing. My only issue is this. It comes w 2 wands and they are both singles. The single is great for stirring things up. It does not make a lot of froth and for some things it’s perfect but sometimes I like a lot of froth and I wish it either came w a single and a double or you could purchase a double separately to intermix. I don’t want to buy a whole other unit just for the double. I wrote to them bc the double frother they sell doesn’t look as good as this one but sometimes a girl needs a lot of foam and sometimes th stirring is perfect.
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Reviewed in the United States on February 20, 2026