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Description
Rat 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 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 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 | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Cyclin Dependent Kinase 5 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| 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. |
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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, cell lysates, and other biological fluids |
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4.1 ★★★★★
Based on 19 reviews
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Product Reviews
★★★★★ 3
The Graphics dont last long
I loved this shirt but after just 1 year, the graphics are gone..
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on August 29, 2025
★★★★★ 5
Everything I needed and more
Color: White
I honestly think this may the first time I ever reviewed a product but I felt like I had to because this thing absolutely ROCKS. So easy to use, so fast, and affordable!!! Finally getting back into photography again thanks to this little thing.
I wasn't sure if it was going to work to be completely honest, but it works and it works great! You can connect it straight to your iPhone, plug in the SD card, go into your files app, and all your pics from the SD card will be there ready for you to choose which you want to download to your photos. Truly everything I needed and more!!!!
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Reviewed in the United States on May 28, 2026
★★★★★ 5
It really works… just one gotcha to be aware of…
Color: White
This little thing is the bomb! I compete in several dog sports with my two dogs, Punk and Shout! I video all our runs with a 4K action camera (GoPro knock-off). The app doesn’t work with my camera, so the only way to scroll through the videos to delete files I don’t need is on the camera itself. It’s very time consuming to watch each video (1-5 minutes long) to see if I need to keep it or delete it. And there is no way for me to process the videos on my phone to post them to FB while at the competitions. So when I found this gadget, I was super stoked. We had a very big 3 day agility competition last weekend, and I was able to use this for the first time. I could easily identify the videos I wanted, download them to my phone, make movies with iMovie, and post them to FB to brag on my dogs. SO VERY HAPPY with the ease of use and reliability. It worked each time I connected it to my phone, it read the micro SD card consistently, it downloaded the videos as selected.
I will warn you — do not use the files app your phone to delete videos from your card. Use the camera to do that. I tested it beforehand and only verified that the videos didn’t show up when I put the card back into the camera, assuming that the videos were actually deleted. Unfortunately, when I did this over the weekend, even though the videos seemed to be deleted, the file space on the SD card was not released. So, my 128G SD card filled up even though I only had 30 1G videos visible on the card. I missed filming one very important event because the camera shut off because the card was “full”. I had to use a different SD card (lower quality) for the rest of the trial. When I got home, the only videos I could recover from the card were the 30 videos, yet the card was still full, even after deleting those 40 files using my PC, the properties of the card still show it full of phantom videos. The only way I could recover all storage space was to re-format the card. So… just beware when deleting files using the Files app on your phone, it does not free up the space on the card EVER.
As a backup plan, I ordered 2 more high-speed Micro SD cards, so I should not run out of space for another 3 day weekend. Overall, I love this little gizmo and will continue to use it A LOT!
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Reviewed in the United States on March 11, 2025
★★★★★ 5
Good SD card reader
Color: White
I needed it for my cameras sd card and it works reammy well, it shows the files really fast, you dont need any instructions to see your files and the design is exactly like on the picture
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Reviewed in the United States on May 13, 2026
★★★★★ 5
Very good
Color: White, Color: White
I have an iPhone 8 and it worked well with this. It was very simple to use. I was able to get all my pictures from my trip and the card fit in nicely as well. I would recommend this and I think the price is right. I was able to get the images right away, it was very fast.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 28, 2026
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