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Description
Mouse Nestin ELISA KitProduct Specification Usage Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High precision pipette and gun tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37 constant temperature box 4. Distilled water or deionized water Sample processing and requirements: Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4 overnight, then centrifuge at 1000g for 20
Product Specification
| Usage |
Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High-precision pipette and gun tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37℃ constant temperature box 4. Distilled water or deionized water Sample processing and requirements: Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4℃ overnight, then centrifuge at 1000×g for 20 minutes, and take the supernatant, or store the supernatant at -20℃ or -80℃, but avoid repeated freezing and thawing. Plasma: Collect the specimen using EDTA or heparin as an anticoagulant and centrifuge at 1000 × g for 15 minutes at 2-8°C within 30 minutes of collection. The supernatant can be assayed or stored at -20°C or -80°C, but avoid repeated freezing and thawing. Cell Supernatant: 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., 1 g of tissue sample to 9 mL 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 the tissue cells, the homogenate can be sonicated or repeatedly frozen and thawed. Finally, centrifuge the homogenate at 5000 × g for 5-10 minutes, and the supernatant can be assayed. Preparation before testing: 1. Please remove the test kit from the refrigerator 10 minutes in advance and equilibrate to room temperature. 2. Preparation of gradient standard working solution: Add 1mL of universal diluent to the lyophilized standard, let it stand for 15 minutes to completely dissolve, and then gently mix (concentration is 10ng/mL). Then dilute to the following concentrations: 10ng/mL, 5ng/mL, 2.5ng/mL, 1.25ng/mL, 0.625ng/mL, 0.3125ng/mL, 0.15625ng/mL, and 0ng/mL. Serial dilution method: Take 7 EP tubes and add 500uL of universal diluent to each tube. 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 step to mix thoroughly. 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 Nestin capture antibody. After incubation and washing, the sample 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 Nestin in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Nestin ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Nestin is a protein encoded by the NES gene. Nestin is a type VI intermediate filament (IF) protein. These IF proteins are primarily expressed in neurons and are involved in radial axonal growth. Seven genes encode the heavy neurofilament (NF-H), medium neurofilament (NF-M), and light neurofilament (NF-L) proteins: nestin and α-internexin in neurons, synemin α and desmuslin/synemin β (two alternative transcripts of the DMN gene) in muscle cells, and syncoilin (also in muscle cells). Most members of this group prefer to associate in tissues as heteropolymers. Nestin forms homodimers and homotetramers in vitro but does not form IFs on its own. In mixtures, nestin preferentially co-assembles with purified vimentin or the type IV IF protein internexin to form heteromeric coiled molecules. | |||||||||||||||||||||||||||||||||
| 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 | Serum, plasma, cell supernatant, tissue homogenate, etc. |
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4.7 ★★★★★
Based on 11 reviews
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Product Reviews
★★★★★ 5
Perfect when you need length.
Color: Black, Size: 15FT Flat Plug
Works great, the ports are perfect, so no more cubes.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 5, 2026
★★★★★ 4
Nice strip, wish it had a power on indicator
Color: Black, Size: 6FT Flat Plug
My first strip was DOA, and seemed to have been returned to Amazon. I say this because I was given a replacement and the packaging was different. First it was shrink wrapped, the first device I received was not. Second, the internal packaging was different.
I scrapped my original review, and this review is based upon the working strip.
This is an attractive power strip with 4 USB charger ports. I like the wall mount holes on both ends that provide a more secure means of attaching the strip to something. Most chargers simply have holes on the back to hang over a screw, this one you can actually fasten to something.
The position of the outlets is good for clunky older power blocks provided they aren't too wide. They don't come close to interfering with the usb ports, ensuring you can easily plug in 4 usb cords and 4 power cords without having them get tangled up.
My chief dislike are the lack of any power indicator. Almost every power strip I have owned in the last 30 years had some visible way to tell the strip was turned on. The other thing is the indicator to show which side of the power switch is on. The off side has a very visible white dot, which would probably make most people believe that was the "ON" position, it is actually the off position. The on position is marked by a raised black bar(or perhaps it is a 1) which is difficult to see. I think I would have just painted the traditional "on" icon next to the power on end of the switch. Again, a power indicator here would have made all the difference.
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Reviewed in the United States on March 14, 2018
★★★★★ 5
terrific, low cost power management tool
Color: Black, Size: 6FT Flat Plug, Color: Black, Size: 6FT Flat Plug
I noticed that most of the power outlets I use on a regular basis are behind large pieces of furniture. Every modern device requires at least a little power (even if only at intervals) and many of them have to be periodically unplugged to be rebooted. This invariably leads to a bulky power strip/surge protector at every usage location.
Most power strip/surge protectors are very bulky and made to sit on the floor, often in a difficult location for routine access. Unfortunately, “unplugging” from a power strip requires holding the strip firmly in place while simultaneously pulling on the plug. With the strip in a difficult location and the fact that it must be restrained to remove the plug, I typically need a scotch to even attempt it.
If that weren't bad enough, most devices use an AC adapter of various different sizes, shapes and orientations. The differences are odd enough that they look like a heterogeneous collection of "rugrats". This further magnifies the problem because power strips are never devised for the space requirements of rugrats. Most power strips assume that everything is a standard 2 or 3 prong plug, lined up in military fashion. So, to make a power strip work, it must often be combined with a variety of cube taps, shifting the vertical dimensions of many outlets to accommodate the different rat dimensions. In some cases, power strips start looking like tenement slums. Power strips that attempt to accommodate the dimensions of rugrats are even bulkier and more difficult to use.
The overall problem was difficult enough that I recently decided to just sacrifice the furniture.
This surge protector power strip has a very low profile with two screw slots at either end. There are locations on all outlet-blocking furniture where the strips can be screw-attached (see photo). This solves nearly all the problems: (a) the strip is convenient to someone seated in the chair or laying on the bed; (b) it’s strongly secured meaning the plugs can be easily removed and replaced with one hand; (c) it has one wide-spaced outlet for the odd rugrat; and (d) it has multiple USB ports with one ‘C’ type charging port, thus eliminating one rugrat.
Given all that and that fact that it’s very reasonably priced, I couldn’t be happier.
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Reviewed in the United States on September 1, 2023
★★★★★ 5
Best Power strip to organize all your electronics.
Color: Black, Size: 6FT Rotating Plug, Color: Black, Size: 6FT Rotating Plug
The best extension cord I ever had so far, it's my 1st day using it but my life has changed for the better. I order yesterday they arrived today, very quick delivery. They were packaged nice and neat with instructions and screws if you want to mount. I love the Rotating Flat Plug, very nice and neat. My life is very busy and I need to be organized with 2 cell phone, laptop, smart watch and headphones. I was having to charge one or two at the time with plugs all over the place. Then having to take charger back in forth from my office to my bedroom and to work. Too much of a headache, I been looking all week and I needed 3 port c and 1 usb. Game changer for the best and I highly recommend to buy. I bought 2 and will be buying 2 more. I bought black to match my furniture. The best $20 I ever spent!!
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Reviewed in the United States on February 15, 2026
★★★★★ 5
Multiplug multiuse
Color: Black, Size: 6FT Rotating Plug
Very functional I like
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 7, 2026
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