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Description
Rat NSMASE 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: 1. 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
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: 1. 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. 2. 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. 3. 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 sphingomyelin phosphodiesterase 2 (NSMASE) capture antibody. After incubation and washing, the assay 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 sphingomyelin phosphodiesterase 2 (NSMASE) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Sphingomyelin phosphodiesterase 2 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Neutral sphingomyelinase (NSMASE), also known as sphingomyelin phosphodiesterase 2 (SMPD2), is an enzyme encoded by the SMPD2 gene. This gene encodes a protein that was initially identified as a sphingomyelinase based on sequence similarity between bacterial sphingomyelinases and yeast proteins. Subsequent studies have shown that its biological function is unlikely to be a sphingomyelinase, but rather a lysophospholipase. | |||||||||||||||||||||||||||||||||
| 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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4.4 ★★★★★
Based on 5 reviews
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Product Reviews
★★★★★ 5
I L-O-V-E these!!!!
Size: 9 Women/8 Men, Color: 1a-407 White
UPDATE: I've tried several difference sellers/brands and keep coming back to these Seekway shoes. I think the key is for me that they are one-piece like water socks with ties rather than having separate tongues. Some other brands, the tongues are long and rub against your foot. These also have a very thin heel cup unlike others that were fine for short term, but on long walks, the thicker heeled ones rubbed nasty blisters in both heels. These Seekway DID NOT. The only down side is that the soles are very thin, so even with orthotics (Happy Step memory foam), when walking for several miles, I did find the balls of my feet sore. But in time, they've actually "toughened up" and now I walk about 3 miles per night in these shoes without any issue. Just bought my third pair!
These shoes are soooooo comfortable with great tread and very breathable. I LOVE these! The price is very reasonable while quality is good. I have one foot that is slightly narrower, especially in the heel, than the other. After a bit of experimenting with the elastic ties, they fit great. True to size. And they look up-scale... unlike some old beach shoes I have from a few years ago. I actually forgot I had shoes on, that's how comfy they are. These are not for support, but that's not their purpose. If you like walking around barefoot yet want protection from injury or stepping on something icky, these are perfect. I also hear some podiatrists are praising barefoot-style shoes as good for building muscles in the foot and legs. Total win!
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Reviewed in the United States on February 24, 2026
★★★★★ 4
Good fit and comfort.
Size: 11.5 Women/10.5 Men, Color: 1o-506 Black
These fit well and are easy to wear in the water. They drain and dry quickly so you can move from wet to dry without having to change shoes. They kept my feet reasonably warm on a chilly day in the river. However the grip on the soles could be a bit better; they got pretty slippery when walking on rocks. They also don't offer much protection if you happen to step on something sharp. They're good if you need something versatile but I wouldn't recommend for serious river work.
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Reviewed in the United States on May 22, 2026
★★★★★ 5
Perfect for Cenotes – Lightweight, Quick-Drying & Comfortable
Size: 6 Women/5 Men, Color: 1a-407 White, Size: 6 Women/5 Men, Color: 1a-407 White
Perfect water shoes! The material is thin, but they dry very fast, which was exactly what I needed. I’m usually a size 6.5 and sized down half a size, and the fit was perfect. I used them for swimming and walking in cenotes in Mexico for about two and a half hours, and they were comfortable the entire time. Great grip and very practical!
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Reviewed in the United States on April 4, 2026
★★★★★ 5
Great quality & comfortable all day wear
Size: 13.5 Women/12.5 Men, Color: 1e-414 Gradient Blue 35
I purchased these for my husband to wear on our vacation to Jamaica. I planned a trip to one of the many beautiful falls and water caves" in the area. The travel instructions stated we would be "climbing rocks & walking through fast moving water" and water shoes with good traction were advisable. The excursion was amazing and my husband was very impressed with the quality and durability of these water shoes. They kept his footing steady, kept him from slipping and were extremely comfortable for all-day day wear. We strongly endorse these water shoes for any water sport or easy summer wear.
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Reviewed in the United States on April 30, 2026
★★★★★ 5
I would give these 6 stars if I could
Size: 7 Women/6 Men, Color: 1a-407 White, Size: 7 Women/6 Men, Color: 1a-407 White
We're going on a trip soon where we'll be jumping into some natural waterfalls and it is recommended that we wear water shoes. I've had water shoes before and so far not a fan. They're tight and ugly, making my feet look like a squishmallow (like rock climbing shoes).
Searched on Amazon and saw these. Hey! The pics online actually looked pretty neat. They reminded me of those funny shoes with toes. I read the reviews and everyone said they were the most comfortable shoes ever. So, I drank the koolaid and bought a pair for me (white) and a pair for hubby (grey/green). We just tried them on.....
So, I'm normally a size 7 (women's) and my feet are wide. For slippers, I fit into a 6. So I took a chance and ordered size 37 (which is honestly the normal EU size for me, but being that these are made in China, who knows what I'll get). Hubby is normally a size 10 (men's) and his feet are very slightly narrower than normal. So again, we took a chance and ordered him size 42 (besides, the size 9.5 wasn't even available as an option).
Holy mother of metatarsals what sorcery is this? I put these on my feet and they felt like a soft hug! Wow! And they fit! At first I thought...oh no, they're a bit BIG. But I was sitting and so when I stood up, my toes went to the tip of the shoe. Remember how I said my foot is wide? Well oddly enough, I had to pull the straps really tight so that they'd feel more secure. This is the only downside: Because I tightened the straps quite a bit, there's a lot of extra elastic (6-8" lenght) that is going to dangle around the top of my foot (see pic). So a lot of creative tucking it in is necessary. Hubby's shoes fit him perfectly. He didn't have to tighten his as much.
Very impressed by the construction of the soles; they feel sturdy and solid and flexible yet thick enough to prevent sharp rocks from poking through. I also love the shape of the toe box. It's a square shape but flattering (on me as my foot is wide). Plus, because of the mesh fabric, stitching, and detail, the shoes look like stylish athletic shoes and not plain neoprene water shoes like the kids wear. They are also extremely light and have holes in the bottom for drainage. The fabric is super soft and stretchy and they are a snap to put on. Super awesome!
Not gonna lie: I don't even have socks this comfy.
Could I have gone a size down? I probably could but because of my wide feet, that would be too risky. If you are a size 7 naturally with narrow feet, I tend to think yeah...you may want to consider sizing down a half size. But my toes hit the tip of the toe box when standing up and I don't want them to overhang. Any tighter and they may fit but they won't look good imo.
I could see why people want to wear these all the time. They're so comfy, I bet you can even sleep in these.
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Reviewed in the United States on January 3, 2025