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Description
Human NDN 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 ultrasonically disrupted or repeatedly frozen and thawed. Finally, the homogenate is centrifuged at 5000×g for 5-10 minutes and the supernatant is collected 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 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 a necdin (NDN) 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 necdin (NDN) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Necdin ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Necdin, also known as NDN, is a protein encoded by the NDN gene. Located in the Prader-Willi syndrome (PWS) deletion region, it is an imprinted gene expressed only from the paternal allele. The protein encoded by this gene may inhibit postmitotic neuronal growth. It functions to stimulate growth regulation and DNA-dependent transcriptional regulation. Associated disorders include Prader-Willi syndrome and Prader-Willi syndrome due to paternal deletion 15Q11q13 type 1. | |||||||||||||||||||||||||||||||||
| 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 and other biological fluids |
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4.6 ★★★★★
Based on 17 reviews
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Product Reviews
★★★★★ 5
The softest, prettiest pink ever. So finely constructed in every detail.
Size: One Size, Color: Crinkle Morning Beach
I recently ordered this Kate Spade Large Leather Shoulder Bag in the Crinkle Morning Beach color, which is a very soft, mellow, light pink. Think puffy clouds dusted with the pink highlights at sunset kind of light pink. I think it’s absolutely gorgeous!
I love the smooth, simple lines of this bag and the “rocking chair” curved bottom panel. The bottom panel allows this purse to stand upright, making it easy to see into it and access the contents. It has a simple thin shoulder strap tied into a perky loop detail at the side with a little tail. Nothing fussy. Just pure simplicity and class with a flirty little flair.
As mentioned in the description, this is a large bag, 14” wide, perfect for travel or if you have more to carry on that day. It’s quite roomy inside. The zipper opens up along the entire top of the bag, making it easy to see into it and take things in and out of it. There’s an open pocket against one side and a zippered pocket against the other to keep things neatly organized and easy to find.
The construction of this bag is perfect in every detail. The leather is just exquisite. So smooth to the touch. Even the light-colored inner lining is cool and smooth. Every surface has neat topstitching in perfect, small stitches in a matching pink thread color. It has a quality zipper that zips smoothly and has a small leather pull. It seems everything about this shoulder bag provides a delightful experience in its use.
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Reviewed in the United States on May 10, 2026
★★★★★ 5
A beautiful, soft, finely finished and costly bag
Size: One Size, Color: Crinkle Morning Beach, Size: One Size, Color: Crinkle Morning Beach
I defer to Mrs. Engineer in everything related to purses. I’ve included her comments below.
UNPACKING
The bag arrives surrounded by a thin foam pad inside a plastic bag. An unremarkable polyester storage bag (shown in a photo) is included. It comes folded in the same package as the bag.
IMPRESSIONS
The first impression the bag makes is that it's simple and beautiful and that the leather feels wonderful.
The interior has one open pouch and one with a zipper (see photos). That’s sufficient for Mrs. Engineer but some people may want more subdivisions of the interior.
The bag has top stitching everywhere. As shown in photos, there is impeccable top stitching throughout the interior and on the bottom.
The interior is light colored, a feature always desirable in a bag for ease of finding things.
The wide, flat (from front to back) bottom allows the bag to sit upright unless it’s loaded in an extremely unbalanced way. Because of the fineness of the leather, most people wouldn’t want to put it down on any flooring unless it was carpeted.
The handle is comfortable and is suitable for both handheld and over-the-shoulder carrying. The strap length can be adjusted to fit the user.
PRICE
It’s difficult to comment on the price and whether it’s a good value. Certainly all the functions of this bag, including looking elegant, can be accomplished with a bag costing a quarter as much. Whether the quality of the leather, the fineness of the finishing and the cachet of the Kate Spade brand is worth the cost of the bag depends on the eye, and the resources, of the beholder.
We received this bag at no cost through the Amazon Vine program in return for an unbiased review. For us, personally, it would not be worth its price at the time of this review, but the popularity of the brand proves that many others feel differently about it.
CONCLUSIONS
This is a beautiful and finely finished bag that we are happy to have, and there’s nothing negative to say about it. I’ve given it five stars.
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Reviewed in the United States on March 26, 2026
★★★★★ 5
Favorite hat
Size: One Size, Color: Tan - 7 Panel
High quality hat.
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Reviewed in the United States on March 1, 2026
★★★★★ 5
Hat goes on head
Size: One Size, Color: Tan - 7 Panel
Another hat for my son he loves it great fit good colors
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Reviewed in the United States on December 27, 2025
★★★★★ 5
One of the nicest hats I’ve bought
Size: One Size, Color: Timber
One of the nicest hats I’ve bought, comfortable, looks great and doesn’t feel cheap.
Different would order again
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Reviewed in the United States on October 23, 2025