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Description
Rat 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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| 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 | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat 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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4.1 ★★★★★
Based on 19 reviews
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Product Reviews
★★★★★ 5
Prefect for our trip.
Color: Muted Pink+Lilac, Size: Large
I bought this travel holder because of the strap that I can fit around my neck. The rings and clasps are metal. There are two rings on the back side to hook the strap to which I really like because less tension on the neck. There was plenty of room to fit our passports with protective covers into the slots on the right. I adjusted the strap long enough that it still lets me easily un-zip the holder while keeping it securely around my neck. No worries about accidentally leaving it somewhere. Only time can tell how much it will hold up to ware and tare. Feels and seems it will hold up.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 3, 2026
★★★★★ 5
From travel chaos to an efficient and methodical process
Color: Haze Blue+Dusty Blue, Size: Large
This organizer was meticulously developed for holding passports, IDs, credit cards, money, tickets, ink pen, car key, and etc. in a compact, convenient, and orderly manner. I also placed Lysol wipes in the exterior front pocket and zipped my cell phone into the exterior back pocket. It is easy to open and retrieve the items needed for every step of my travels. No more digging for travel essentials in a variety of zipped pockets. Travel chaos is now an efficient and methodical process.
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Reviewed in the United States on February 19, 2026
★★★★★ 5
Exelente.
Color: Gunmetal Grey, Size: Large
Exelente producto estoy muy conforme.
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Reviewed in the United States on May 5, 2026
★★★★★ 4
Light, organized, security concern about strap
Color: Black+Gunmetal Grey, Size: Large, Color: Black+Gunmetal Grey, Size: Large
I used this passport holder on a recent multi passport trip and overall it does what it claims without feeling bulky. The nylon material is lightweight but reasonably sturdy and holds its shape well in a backpack or day bag. It does not feel flimsy, and still looks new with the stitching and zippers holding up well
Organization is the strong point. On one side I was able to fit three passports comfortably, though the pockets are not a uniform depth. The pockets are loose enough you can also double up if needed. A fourth passport fits on the opposite side along with several credit cards. There are zippered pouches inside and outside useful for cash and a phone, plus open access pockets that work well for boarding passes or documents you need to grab quickly. There is also a small storage area above the credit card slots that turned out to be useful, though listed for SSD storage, I stored a couple of CR2032 batteries there for Bluetooth trackers.
The RFID blocking is a basic but expected feature and worked as intended. Nothing about the layout forces cards or passports to bend, which I appreciate since I tend to keep this packed while moving through airports.
The shoulder strap is my main criticism. It is thin and functional but could be cut fairly easily. I would prefer to see embedded wire or a more reinforced strap for better security in crowded areas. The handle is fine for short carries, but the strap is what you rely on most of the time.
The included Bluetooth tracker works well enough for proximity alerts, but it is not compatible with Apple Find My. I ended up removing it and using an AirTag instead, which fit without issue and gave better ecosystem integration for me.
Overall, this is a practical, lightweight travel organizer that handles multiple passports and documents well. The layout is thoughtful and flexible, and the build quality matches the price point. The strap and tracker limitations are worth noting, but the core function as a family passport holder is solid.
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Reviewed in the United States on January 11, 2026
★★★★★ 3
Nice for travel
Color: Black, Size: Large
No fashion statement here but useable for my trip.
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Reviewed in the United States on May 8, 2026
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