Shipping Estimate
USA
- USA
- CAN
- USA
- CAN
Ships within 48 hours · Estimated delivery Jul 7 - Jul 12
Promo Codes Available:
For Your Every Summer RSVP, with Code: SUMMER15
Description
Human CDK5 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. |
|||||||||||||||||||||||||||||||||
| 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 cyclin-dependent kinase 5 (CDK5) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by HRP peroxidase and to yellow by acid. The intensity of the color is positively correlated with the amount of cyclin-dependent kinase 5 (CDK5) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Cyclin Dependent Kinase 5 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
|
|||||||||||||||||||||||||||||||||
| Background | Cyclin-dependent kinase 5 (CDK5) is a protein, or more specifically, an enzyme, encoded by the Cdk5 gene. It is prominently expressed in postmitotic neurons of the central nervous system (CNS). This molecule belongs to the cyclin-dependent kinase family. It was originally named NCLK (Neuronal CDC2-Like Kinase) because it shares a similar phosphorylation pattern. It is also known as Tau protein kinase II when it binds to activator kinases. Furthermore, Cdk5 is involved in T cell activation and plays a crucial role in the development of autoimmune diseases such as multiple sclerosis. | |||||||||||||||||||||||||||||||||
| 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. |
|||||||||||||||||||||||||||||||||
| 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 |
Shipping Notes
- Free Standard Shipping on $100+ Orders to the USA.
- Except Preorder products are shipped in 48 hours.
- Delivery to the USA:
- Standard Shipping : 3-10 business days
- If time is of the essence, please consider selecting expedited delivery for faster service.
Exchange/Return Notes
- We offer a 30-day return/exchange service after receiving.
- Final sale items are not eligible for returns or exchanges.
- To process your return/exchange, please contact us at [email protected]
- Please click here for more details>>> Return & Exchange Policy
4.7 ★★★★★
Based on 357 reviews
Sort
Product Reviews
★★★★★ 5
**Reliable and Efficient: Nilight 50FT 14AWG Speaker Wire Review**
Size: 14AWG-50FT
I recently set up my home theater system using the Nilight 50FT 14AWG Copper Clad Aluminum Wire, and I'm quite satisfied with its performance. The 14 gauge thickness is ideal for ensuring high-quality sound transmission, which is crucial for a setup like a 5.1 surround sound system. Although it's not pure copper, which some audiophiles might prefer, the copper clad aluminum (CCA) used in this wire provides a great balance of quality and value. It's flexible, lightweight, and offers good corrosion resistance, making it an excellent alternative to more expensive pure copper wires.
The wire comes in a 50-foot length, which was just enough for my setup with speakers placed at various points around the room. One of the features I appreciate most about this product is the color-coded PVC jacket. It simplifies the installation process by making it easy to differentiate polarity, reducing the risk of errors during setup. The wire is also easy to handle; it cuts and strips smoothly, allowing for a clean and secure connection.
The environmental PVC jacket provides good elasticity and fire resistance, adding an extra layer of safety to its use in home environments. While the jacket is thick, it's important to note that it isn't shielded, so I made sure to route it away from power cords to avoid potential interference.
Overall, the Nilight 14AWG wire is an excellent choice for anyone looking to install a reliable and effective speaker system at home. It's straightforward to use, provides great performance, and comes at a cost-effective price point. I would definitely consider purchasing it again for future projects. Whether you're setting up a new system or upgrading an existing one, this wire is definitely worth considering.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 29, 2024
★★★★★ 4
Great for the price!
Size: 14AWG-50FT, Size: 14AWG-50FT
Ordered a spool of wire to install a new light bar and rock lights on my Jeep. Did the trick and cheaper than most other brands.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on February 17, 2026
★★★★★ 5
Decent copper clad aluminum
Size: 14AWG-50FT
If you want the best wire, you should probably go with pure copper wire. It is more expensive but conducts better and thus is more efficient and less likely to heat up. If you are doing a low power, short run of cable, however, this wire should serve your purposes well.
My purposes for such wire are varied. I am a licensed ham, like to build circuits, and have an electrical engineering degree. Depending on the specific tasks I'm on, some I'll gladly use this wire, others I'll go with pure copper.
In comparison to other similar copper clad aluminum wire I've used in the past, the wire is pretty flexible and easy to work with. I also prefer the softer insulation like this has. You need to fit the wire to the job but if you have the job this is right for, it'll serve you well.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 22, 2024
★★★★★ 5
good wire for the price
Size: 14AWG-50FT
This speaker wire is of good quality. I don’t know what to say about it besides that its 14 gauge, which is a bit on the thick side, but good for getting high quality sound or for doing long runs of cable. Its not pure copper, but that's not really necessary unless you are an audiophile and even then, i wonder if its really noticeable. The 50 feet it comes with is enough for a 5.1 surround system, as long as your speakers aren’t too far away. I haven't noticed any weird sounds from my system after setting it up with this cable. The housing is relatively thick but not shielded, so I was careful to keep it away from power cords. All in all this is good cable, especially for the price, and I would purchase it again.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 23, 2024
★★★★★ 4
Not as good as pure copper, but maybe good enough
Size: 14AWG-50FT, Size: 14AWG-50FT
This wire is copper-clad aluminum, which means you sacrifice some conductivity for a for a less expensive conductor. This wire is very lightly clad with copper - the thinness of the copper coating is obvious from a casual observation (see image).
I measured 4-wire resistance across the entire length (50') of wire - it comes out to 0.34 ohms on each lead. This is a bit less than double what you'd expect from an equivalent length of the same gauge of multistrand copper, so if you're planning on running a long length of this it might be prudent to pretend that you're working with a slightly smaller gauge than you actually are. That is exactly the approach I'm taking with it, and it works well for me within that context.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 19, 2024