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Description
Human NELL2 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 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 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 capture antibody against protein kinase C-binding protein NELL2 (NELL2). 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 protein kinase C-binding protein NELL2 (NELL2) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Protein kinase C-binding protein NELL2 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | NEL-like protein 2, also known as NELL2, is an enzyme encoded by the NELL2 gene. This gene encodes a cytoplasmic protein containing epidermal growth factor (EGF)-like repeats. The encoded heterotrimeric protein may be involved in cell growth regulation and differentiation. Several variants exist, encoding several different isoforms, at least one of which appears to be a secreted protein. Research suggests that this protein plays a role in neuronal cell growth and differentiation, as well as tumorigenesis. It is involved in regulating hypothalamic GNRH secretion and controlling puberty. | |||||||||||||||||||||||||||||||||
| 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, and other biological fluids |
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4.3 ★★★★★
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Product Reviews
★★★★★ 5
My favorite Melatonin supplement
Size: 100 Count (Pack of 1), Style: Melatonin 1mg
I have used this for a few months and really like it. I had previously used a spray that for some reason had added B6 (>2000% RDA) and I developed toxic levels of Vitamin B6 so had to switch. I love that this doesn't have any added garbage including no added flavors, artificial sweeteners or artificial dyes. The taste is fine. It's not candy and isn't supposed to taste like candy. It is just a bland taste that I hardly notice. Dissolves quickly and easily. Love it.
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Reviewed in the United States on September 27, 2024
★★★★★ 5
TLDR: Don't wait! Buy it now! This stuff is really great and I think you're gonna love it!
Size: 90 Count (Pack of 1)
Life Extension Neuro-mag:
I first learned about magnesium l-threonate on YouTube. It was recommended by several physicians there and in person, all of whom consistently make good suggestions in terms of health optimization. Even still, I didn't go for it right away, unsure if this form of magnesium could really be much different than or better than the liquid magnesium from Life Extension that I've been using with great results for several years now. Still, I was curious, as I did some more research and I kept hearing and reading that magnesium l-threonate is “different” because it crosses the blood-brain barrier. Now I'm not gonna try to explain in detail what that is, but I had an understanding of it enough that hearing this further intrigued me. It's my understanding that the human body has this protective measure in place to protect us and for very good reasons so I wanted to be sure I'm making a safe decision for me. As an overview, basically you want the good stuff you consume, that your brain needs to function, to be able to make it to your brain- but not everything. I don't know how our bodies work this out, but apparently they do, so we don't have to figure it out on our own. Imagine that, trying to tell your gut where to send each calorie and nutrient etc.? Anyway, I decided to give this stuff a chance and within 3 days I knew that I will never go without this form of magnesium again if I can help it. For me, it's really that amazing, and of course as a consumer, I hesitate to say, but it's absolutely worth the asking price and more. I've had years of experience with this brand now too and it's been top notch from the packaging to the product, so I will keep buying from Life Extension. With this I sleep better, problem solving is less stressful and my energy level has improved. It does have the relaxing effect on my body that other forms of magnesium do too, but this one has such a profound effect on my mind it's very difficult to quantify, or put into words in text that can even begin to convey or actually help you comprehend just how powerful the effects are. Kind of like the old saying “you had to be there” I suppose. That said, eventually I did buy the magnesium l-threonate from Life Extension and it is quite literally one of the best decisions I've EVER made in my nearly 50 years. It's weird to say that, am I really that old? Seriously, if you do some research and find that the effects sound like the type you might benefit from- buy it!
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Reviewed in the United States on February 24, 2024
★★★★★ 5
Great Product
Size: 90 Count (Pack of 1)
Life Extension’s Neuro-Mag Magnesium L-Threonate is a standout supplement designed to support cognitive health, and after trying it for several months, I can see why it’s so highly regarded. This product contains magnesium L-threonate, a unique form of magnesium that’s specifically formulated to cross the blood-brain barrier, making it highly effective for brain health. Each serving (three vegetarian capsules or one scoop of the tropical punch powder) delivers 2,000 mg of magnesium L-threonate, providing 144 mg of elemental magnesium. Here’s my take on its performance, benefits, and overall experience.
Pros:
• Cognitive Support: I noticed improved mental clarity and focus after about two weeks of consistent use. Tasks that require working memory, like recalling details or multitasking, felt smoother and less taxing. This aligns with research indicating magnesium L-threonate’s ability to enhance synaptic density and cognitive function, particularly in aging individuals.
• Sleep Benefits: While not marketed primarily for sleep, I found that taking it about an hour before bed helped me fall asleep faster and wake up feeling refreshed. This echoes user experiences shared on platforms like X, where some reported better sleep quality and daytime alertness.
• High Absorption: Unlike other magnesium forms like oxide or citrate, which are better for whole-body benefits, magnesium L-threonate is designed for brain absorption. Studies cited by Life Extension show it can boost magnesium levels in cerebrospinal fluid by 15% in animal models, which likely contributes to its cognitive effects.
• Clean Formula: The capsules are gluten-free, non-GMO, and vegetarian, with no major allergens or unnecessary fillers. The powder version has a pleasant tropical punch flavor, making it easy to mix with water or juice.
• Trusted Brand: Life Extension’s rigorous testing for purity and potency gives confidence in the product’s quality. Their transparency, including offering Certificates of Analysis, is reassuring.
Cons:
• Price Point: Neuro-Mag is pricier than standard magnesium supplements, which might be a drawback for budget-conscious buyers. However, the specialized form and targeted benefits justify the cost for those prioritizing brain health.
• Time to Notice Effects: While some users report benefits in as little as six weeks, full effects may take up to 12 weeks, requiring patience.
Final Verdict: Life Extension’s Neuro-Mag Magnesium L-Threonate is a high-quality supplement that delivers on its promise of supporting cognitive health. Its ability to enhance memory, focus, and even sleep quality makes it a worthwhile investment for those prioritizing brain function. Great benefits and clean formulation make it a top choice!
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Reviewed in the United States on June 16, 2025
★★★★★ 5
Effective results
Size: 90 Count (Pack of 1)
This is the only supplement I take where I actually feel its effects. It helps regulate my sleep, helps brain function and anxiety, just makes me feel better overall! And it’s not rough on the stomach at all. Highly recommend!
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Reviewed in the United States on May 19, 2026
★★★★★ 4
Noticeable Support for Calm and Clarity
Size: 90 Count (Pack of 1), Size: 90 Count (Pack of 1)
This magnesium L‑threonate supplement has been a great addition to my daily routine. The capsules are easy to take, gentle on my stomach, and I’ve noticed a clearer, more focused feeling throughout the day. It also brings a nice sense of calm in the evenings without making me groggy. The formula feels clean and well made, and it fits effortlessly into a wellness routine. If you’re looking for a reliable magnesium option that supports relaxation and mental clarity, this one is a solid choice.
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Reviewed in the United States on January 29, 2026