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Description
Human NLRP3 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 sonicated or repeatedly frozen and thawed. Finally, centrifuge the homogenate at 5000×g for 5-10 minutes, and collect the supernatant for analysis. Cell Lysis Buffer: Gently wash adherent cells with pre-chilled PBS, then trypsinize and collect the cells by centrifugation at 1000×g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash the collected cells three times with pre-chilled PBS and resuspend 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 freezing and thawing or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and collect the supernatant for analysis. Cell Culture Supernatant: Centrifuge at 1000×g for 20 minutes. Collect the supernatant for analysis, or store at -20°C or -80°C, but avoid repeated freezing and thawing. 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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| Sensitivity | 0.18 ng/mL | |||||||||||||||||||||||||||||||||
| 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 an NLR Family, Pyrin Domain Containing Protein 3 (NLRP3) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB converts to blue under the catalysis of HRP peroxidase and to yellow under the action of acid. The intensity of the color is positively correlated with the amount of NLR Family, Pyrin Domain Containing Protein 3 (NLRP3) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human NLR Family, Pyrin Domain Containing Protein 3 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | NLR family pyrin domain-containing 3 (NLRP3) (formerly known as NACHT, LRR, and PYD domain-containing protein 3 [NALP3] and cryopyrin) is a protein encoded by the NLRP3 gene located on the long arm of chromosome 1. NLRP3 is primarily expressed in macrophages and, as a component of the inflammasome,436 detects products of damaged cells, such as extracellular ATP and crystallized uric acid. Activated NLRP3, in turn, triggers an immune response. Mutations in the NLRP3 gene have been associated with several organ-specific autoimmune diseases. | |||||||||||||||||||||||||||||||||
| 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 | 0.312-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates, cell lysates, cell culture supernatants, and other biological fluids |
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4.4 ★★★★★
Based on 1882 reviews
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Product Reviews
★★★★★ 5
Exceeded expectations in every way.
Size: 2' x 3' (Sheepskin shape), Color: Gray Tip, Size: 2' x 3' (Sheepskin shape), Color: Gray Tip
I purchased a pair of these a week apart, because the price seemed too good to be true. Instead, they have exceeded my expectations. I own a Buick with leather seats that look incredible, but are not comfortable on long trips, and my wife and I were preparing to leave on a 2200 mile round trip vacation. On the first day, we drove 700 miles, much of it through desert. We were tired when we reached our destination, but comfortable and cool for every mile of it. The sheepskins have made such a difference, that I have never removed them from the car. It is now July, with triple digit temperatures every day, surrounded by desert. The sheepskin keep us cool on every drive. I could have purchased sheepskin seat covers, but these provide the same comfort for a fraction of the cost. Yes, it takes a small amount of effort to adjust them when you first get into the car, but I don't even think about it any more. As far as the quality, there is no shedding, no smell, and no discoloration. We have put over 4,000 miles on the car since I put these in, and I have no complaints to register.
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Reviewed in the United States on July 18, 2025
★★★★★ 5
Just buy it!
Color: Grey, Size: 74"x60"
I'm always cold and wanted a blanky, a goto cuddly blanket! I bought this with the intentions of turning it into a duvet for my heating blanket. Now, I haven't bought a blanket in over 20yrs and then it was just some down comforters where I updated the duvets over the years but that's it.
So I started shopping and picked my top 5 faux fur blankets based on fiber density (where available), color, and customer reviews. I was willing to pay as long as I got that goto blanky, I didnt want to be left underwhelmed, especially the more I have to spend.
As a blanket newbie I got stuck and couldn't decide, so it came down to customer reviews and price point. There is $600 brand of blanket which im.sure is lovely but I will never know, just crossed them off my list, couldn't justify it, leaving this as the next most expensive on my list.
So, I asked myself, do you really get what you pay for? Or all these blankets made by the same company, basically the same with different price tags? After reading EVERY SINGLE REVIEW ON ALL 5 BLANKETS and some others I had crossed out because I'm so terrified of buyers remorse, THIS BLANKET HAS THE FEWEST NEGATIVE COMMENTS OF ANY BLANKET!!! That being said theres a lot of postive comments out there but what if the other blankets positive comments are just from people who've never experienced a REALLY NICE BLANKY??? So I went with highest price point willing to spend (under $300ish) and fewest negative comments from people who sounded like they knew blankets, pics and videos people uploaded, etc. I let go and turned it over to the universe...
If you're on the fence because it is pricey, JUST GET IT! It's freaking nice, like REALLY NICE! It's as if not more comfy than my 20 year old ralph Lauren duvet which I spent way more on than this.
THIS BLANKET IS AMAZING and IT DOESNT SHED!
1. I'm not a trapper or a tanner but it looks real af to me. Real enough...
2. The fur is long, soft and fluffy! It's like your friends shaggy dog you want to love and squeeze but you know he hasn't been brushed and can smell him when you walked in the door. It's like that but only positive, I never want to leave it , pet it like dog, it's becoming my best friend, and I don't have to walk it! Even the inside is soft! It's like husky on th outside and rabbit on the inside!!!
3. It's warm and It's got some weight to it! Definitely not a cheap throw!!!!! Remember I was gonna use this as a duvet cover for my electric blanket??? Under this alone I sweat! Several times a night i wake up and throw it off me, my electric blanket doesn't even make me sweat and which is why I thought I could use cover for it.... WRONG! This blanket was all i needed!
I've already probably said too much, but this Is the first review I've ever written. All I can say is if you're undecided;
JUST GET THIS DANG BLANKET, SERIOUSLY!
One more thing, to tell the world how crazy my fear of buyers remorse is a week or so ago, my number 3 blanket, leedy much cheper than this, was on ridiculously low sale so I had to buy it too see I you get what you pay for??? And YOU DO! Granted, It's a really nice blanket, and I probably would have been fine with it, HAVING NEVER SEEN THIS BLANKET.... The other blanket soft, shorter hair, looks real enough, sheds a bit so I brush it, but compared to this blanket, well you can't! It's like comparing a howrd Johnson or days inn to the ritz-carlton, or tube hamburger and a filet, one of these blankets you might consider cutting up for a kids costume and the other you definitely would not and its not just because of the price differnce, but in this instance, you do get what you pay for! The other blanket is not like us, hahaha
If you made it this far I hope this helps you, good luck, and I'm telling you...
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Reviewed in the United States on February 21, 2025
★★★★★ 5
Buy this while it’s priced so low.
Color: Brown, Size: 74"x60"
I am a collector of furry blankets and this one is my new favorite. It’s heavy, and really warm. It’s also a beautiful blanket with the hair tones differently than I’ve seen before. Thank you for the high quality and a relatively lower price than other of close to the same value. It’s so soft and beautiful I had to buy another one!
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Reviewed in the United States on September 30, 2025
★★★★★ 5
Luxury blanket it's everything I expected it to be
This is a nice soft smooth blanket it feels almost like a real animal it's not super heavy but it's a good weight it has a dense short hair best for someone who doesn't want a super furry blanket it's very affordable for the quality it's very warm its shiny in the light I love it I haven't washed it but im sure it would wash easily. it seems durable doesn't feel cheap treat yourself to luxury and get this blanket
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Reviewed in the United States on December 6, 2025
★★★★★ 4
Look extremely good and very good quality feel.
Color: Brown, Size: 74"x90"
Very warm and comfortable. Moreover, feel great.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on December 16, 2025