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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. |
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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 | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human 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 | 0.312-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates, cell lysates, and other biological fluids |
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4.2 ★★★★★
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Product Reviews
★★★★★ 5
Excellent Quality Wire Lugs and Adhesive Heat Shrink Set
Size: 2/0 Awg - 3/8" (M10) Ring, Number of Items: 10
I recently purchased the 10pcs 2/0 AWG-3/8" Wire Lugs Ring Terminal Connectors along with the 10pcs 3:1 Dual Wall Adhesive Heat Shrink set from Amazon and I must say, I am thoroughly impressed with the quality and performance of this product!
Firstly, the wire lugs are exceptionally well-made with heavy-duty materials. The copper construction ensures excellent conductivity, making them ideal for various electrical applications. The 2/0 American Wire Gauge size with a 3/8" ring terminal is perfect for my specific needs, and they fit securely on the cables, providing a reliable and sturdy connection.
The wire lugs come with the corresponding 3:1 dual wall adhesive heat shrink tubes, which are an excellent addition to the set. These heat shrink tubes offer superior insulation and protection, making sure that the connection is secure and insulated against any potential damage or short-circuiting. The adhesive lining within the heat shrink tubes ensures a tight and waterproof seal, further enhancing the reliability of the connection.
I found the installation process to be straightforward and hassle-free. The wire lugs are easy to crimp onto the cables, and the heat shrink tubes shrink perfectly around the connection using a heat gun. The adhesive within the heat shrink tubes bonds effectively, leaving no room for any moisture or contaminants to compromise the connection.
Overall, I highly recommend the 10pcs 2/0 AWG-3/8" Wire Lugs Ring Terminal Connectors and 10pcs 3:1 Dual Wall Adhesive Heat Shrink set. The quality and performance of this product exceeded my expectations. Whether you're working on automotive projects, marine applications, or any electrical wiring projects, this set will undoubtedly fulfill your requirements. It's a fantastic value for the price, providing reliable and secure connections.
I'm extremely satisfied with my purchase, and I will definitely be purchasing more in the future for my upcoming projects. Don't hesitate – give this product a try, and you won't be disappointed!
Disclosure: I received no incentive or compensation for providing this review. My review is based on my personal experience as a customer.
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Reviewed in the United States on December 27, 2023
★★★★★ 5
Two thumbs up
Size: 1/0 Awg - 3/8" (M10) Ring, Number of Items: 4, Size: 1/0 Awg - 3/8" (M10) Ring, Number of Items: 4
Very nice battery lugs. They are not thin junk & they fit. I used a hydraulic crimper which gave me an excellent crimp. Well worth the money.
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Reviewed in the United States on April 17, 2026
★★★★★ 5
Works fine
Size: 6 Awg - 3/8" (M10) Ring, Number of Items: 10
Decent quality
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Reviewed in the United States on May 26, 2026
★★★★★ 5
Item is worth every penny
Size: 4 Awg - 3/8" (M10) Ring, Number of Items: 10
Very well packaged and very good quality shrink tube super thick 😄 item is as prescribed 💯 Thank You
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Reviewed in the United States on April 25, 2026
★★★★★ 4
Good Terminals
Size: 8 Awg - 1/4" (M6) Ring, Number of Items: 10
Used to make radial wire sets for amateur radio. Good connectors with thick metal. Heat shrink is glue lined. Would purchase again.
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Reviewed in the United States on May 1, 2026