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Description
Human NOSTRIN 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 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 Nitric Oxide Synthase Trafficker (NOSTRIN) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by peroxidase (HRP) catalysis and to yellow by acid. The intensity of the color is positively correlated with the amount of Nitric Oxide Synthase Trafficker (NOSTRIN) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Nitric Oxide Synthase Trafficker ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | NOSTRIN is a protein-coding gene. Disorders associated with NOSTRIN include eclampsia and preeclampsia. Pathways involved include activation and regulation of eNOS and metabolism. A key homolog of this gene is CD2AP. NOSTRIN consists of a single polypeptide chain of 506 residues, 58 kDa, with an N-terminal cdc15 domain and a C-terminal SH3 domain. NOSTRIN mRNA is abundant in highly vascularized tissues such as the placenta, kidney, lung, and heart, and NOSTRIN protein is expressed in vascular endothelial cells. Nitric oxide (NO) is a powerful mediator of biological processes such as neurotransmission, inflammation, and vascular homeostasis. NOSTRIN binds to endothelial NO synthase, the enzyme responsible for NO production, and triggers the translocation of ENOS from the plasma membrane to vesicular subcellular compartments, thereby attenuating ENOS-dependent NO production. | |||||||||||||||||||||||||||||||||
| 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 | 0.15-10 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates, cell lysates, and other biological fluids |
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4.2 ★★★★★
Based on 12 reviews
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Product Reviews
★★★★★ 5
My Baby loves his new baby!
Size: Medium, Size: Medium
Finally! A ball my 107 lb. pitty/pointer mix can’t destroy. He has torn up every single “indestructible” toy in a day or so until now. He peels fabric and fur off non-rubber toys. He is a 4-legged shredder. I’ve added pictures of him for a size reference of my dog versus this product. I know I had wanted that when shopping for this item.
This ball is now his beloved “baby.” He goes absolutely insane looking for it, chewing it, chasing it, and even sleeping with it. He won’t rest unless he knows exactly where it is. It is his only toy, and he is good with that. After getting this home, I tried giving him other choices, but he won’t touch them.
I was worried that he would rip it apart by putting his teeth in the holes, but so far, not a scratch. It is soft enough to collapse just enough to keep it from cracking when his jaws crush it but not so soft that he can rip it to sheds. The holes are large enough to prevent a suction or vacuum effect on his tongue or mouth. For his size jaws, it is a perfect fit. Other dogs might have problems with suction or be able to get their teeth in those holes and rip it apart. Who knows?
The older version of the Chuckit squeaky ball was his favorite for the year and a half he had it, but the way they make them now, he peels off the orange triangle in hours flat, the squeaker falls out (choking hazard), and then he rips it apart.
I’m going to stock up on these. I can’t imagine how crazy he will be if they ever stop production or change the materials.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 9, 2019
★★★★★ 5
FANTASTIC PAD HOLDER
Size: 28'' X 34'', Size: 28'' X 34''
Amazon Basics Dog Training Pad Holder 28X34.
I'll start out by saying I wasn't too sure about it, when I got it. It is awkward to put the pads in by yourself if your short as on each end the sides have to come up together and it’s big, however, I've done it five times alone now and it’s doable. There was no smell, it arrived in perfect condition. My husband was home and we set it up together, I walked into the bathroom to get a soiled pad I had keep to put on it for her to know what to do came back and she had already used it and she’s only four months old. I love it love it love it, it’s not too big, but it’s her home too, and we have it in the living room where we spend a lot of time as we’re retired. When she gets a little older, we will probably move it to another room, but for now it’s staying there. After she urinates, it goes down to a very pale yellow color I have been able to use one pad for all day, however, she does have a pad in a under bed tote in our master bathroom. We’ve had a few guests over and if she hadn’t used it they didn’t even know what it was. I highly recommend it. I have been using pads in an under the bed tote for over 10 years and this is a welcome audition. You won’t be sorry if you have little ones that go in the house.
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Reviewed in the United States on January 1, 2026
★★★★★ 5
Very easy to use and clean
Size: 22'' X 22''
Saved my pad training with my puppy. It's firm and easy to clean. It holds the pads well for the most part, but sometimes my dog gets a little crazy and it comes out, but I don't think it could be better. Still give a 5-star!
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Reviewed in the United States on May 3, 2026
★★★★★ 5
Good product
Size: 22'' X 22''
Easy to change out pads.
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Reviewed in the United States on May 15, 2026
★★★★★ 1
Don’t buy
Size: 28'' X 34''
Mine arrived broken and scratched I think I received one that someone had returned. It also doesn’t hold pads that well my 4 lb teacup Yorkie is able to pull the pad off of the pad holder without even ripping the pad.
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Reviewed in the United States on March 26, 2026
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