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Description
Human NLRC5 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: Adherent cells should be gently washed with pre-chilled PBS, then trypsinized and harvested by centrifugation at 1000×g for 5 minutes. Suspension cells can be harvested directly by centrifugation. Collected cells should be washed three times with pre-chilled PBS and resuspended 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. 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 competitive enzyme-linked immunosorbent assay (ELISA). Samples, standards, biotin-labeled antibodies, and HRP enzyme conjugates are added sequentially to microwells pre-coated with universal species NLR family CARD domain containing 5 (NLRC5) antigens. After incubation and washing, the sample is developed with the substrate TMB. TMB is converted to blue under the catalysis of peroxidase (HRP) and to the final yellow under the action of acid. The depth of the color is negatively correlated with the universal species NLR family CARD domain containing 5 (NLRC5) in the sample. The absorbance (OD value) is measured at a wavelength of 450nm using an enzyme reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human NLR family CARD domain containing 5 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | The NLRC5 protein (NLRC5) is encoded by the NLRC5 gene. It is also known as NOD27, NOD4, and CLR16.1. It is an intracellular protein that plays a role in the immune system. It is a pattern recognition receptor and may be involved in innate immunity to viruses by regulating interferon activity. In lymphocytes, NLRC5 localizes to the nucleus and drives MHCI gene expression by occupying the H-2D and H-2K gene promoters. | |||||||||||||||||||||||||||||||||
| 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 | 0.156-10 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates, cell lysates, and other biological fluids |
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4.3 ★★★★★
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Product Reviews
★★★★★ 5
Just what I needed
Color: Black
This keyboard pad has been everything I expected it to be. Soft yet firm enough for my wrists, simple yet elegant design and reasonably priced. At some point when it wears down I do plan to purchase again.
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Reviewed in the United States on April 5, 2026
★★★★★ 5
Comfortable, Simple, Reliable
Size: Tenkeyless, Pattern Name: Keyboard, Size: Tenkeyless, Pattern Name: Keyboard
This is a phenomenal product, and exactly what I wanted. I used to have wrist pain while gaming; I bought this, and then completely forgot that I used to get wrist pain while gaming.
The foam is firm, but comfortable; it’s very supportive, but also flat and consistent. It’s cool, so my wrist never gets sweaty or hot just from using the wrist rest. Its rubberized bottom prevents it from moving at all while gaming. I’ve used it for about a year, and the foam has no indentation at all; in fact, there’s not noticeable wear of any kind.
My one complaint, that I’ve seen in other reviews as well, is that the fabric skin of the wrist rest isn’t quite fit over the foam: there’s some extra material, and it can wrinkle slightly.
Overall, I’m quite satisfied with the product! It’s completely eliminated my wrist pain, and I’ve gotten a year of use out of it with no problems so far. I’d say that’s more than worth the fair price, and I would buy it again without hesitation!
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Reviewed in the United States on April 17, 2026
★★★★★ 5
Very High Quality Memory Gel Foam
Size: Mouse, Pattern Name: Mouse
Firmness is pretty solid, this material is high quality. This isn't that kind of foam that will start loosing it's shape after a year or two, this is a firm gel like substance and it's pretty comfortable.
The color is black as I expected, it's shape is rectangle just like in the pictures, it's appearance is as described.
The only think is that they do use a polyester cover, which does appear as though it may wrinkle over time, and it will likely not outlast the Gel Foam inside. Like I said though the Gel Foam is legit Gel Foam, I have heard these claims before only to find out that it's something far cheaper, that's not this.
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Reviewed in the United States on April 2, 2026
★★★★★ 5
Awesome
Size: Full Size, Pattern Name: Keyboard, Size: Full Size, Pattern Name: Keyboard
If you spend more than a few hours a day at your keyboard, stop what you’re doing and buy this wrist rest immediately. I was starting to feel serious strain after long typing sessions, and this simple accessory has completely eliminated the discomfort. It’s a true game changer for desk ergonomics.
The material quality is phenomenal. The memory foam (or gel) is perfectly supportive—it’s soft enough to cushion my wrists but firm enough that it doesn't flatten out or lose its shape, even after weeks of heavy use. The non-slip base is also fantastic, keeping it anchored exactly where I need it, no matter how intense my typing or gaming gets.
What I appreciate most is the immediate relief. The height and angle are spot on, positioning my hands and wrists in a neutral, comfortable alignment. The reduction in pressure on my carpal tunnel area was noticeable within minutes. I can now work for extended periods without the usual aches or stiffness.
This is a simple, high-impact product that vastly improves quality of life at the computer. It’s durable, comfortable, and absolutely essential for wrist health. Worth every penny for the long-term comfort it provides!
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Reviewed in the United States on November 7, 2025
★★★★★ 4
Pair with HyperX Keyboard Rest to complete the look!
Size: Mouse, Pattern Name: Mouse
Soft, comfortable, doesn't slide around, even on my XL mouse pad. Cool to touch which is nice.
After a year of use, fabric on top becomes slightly loose and bunches up when moving wrist around.
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Reviewed in the United States on February 18, 2026