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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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★★★★★ 5
BUY THEM NOW!! the search is OVER! 🥳
Color: Blue, Size: Queen Size Set of 2, Color: Blue, Size: Queen Size Set of 2
pillow whore here 🤭 and I very rarely find anything worth writing a 5 star review for but let me just save you the time of searching anymore for the PERFECT pillow!!! BUY THEM! 😍 I promise you will NOT be disappointed!! I have spend thousands on pillows over the years, trying to find the perfect pillows... As someone who suffers from debilitating chronic pain and who has had multiple neck and back surgeries I'm very particular about pillows & have such a hard time finding one's that are comfortable, not too hard, not too soft but also offer great support. I FINALLY found them 🥹 and my hubby is Italian and stays hot but gets even hotter when he sleeps so finding an authentic cooling pillow has seemed impossible UNTIL now!! Just wow!! Now Im adding more to my cart while they are still 70% off 🥳🤩
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Reviewed in the United States on May 8, 2026
★★★★★ 3
no thank you
Color: Blue, Size: Queen Size Set of 2
while they look great, they are not soft. I can not sleep with them. Woke with a headache.
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Reviewed in the United States on May 7, 2026
★★★★★ 5
Loved the pillows
Color: Blue, Size: Queen Size Set of 2
I don’t get paid for reviews so if you want a real one here ya go ….
These pillows are great in my opinion. They came vacuum sealed and popped right up “ big and fluffy” immediately, as in big enough I wasn’t sure they would fit in my pillow cases.
I used them last night and even though they are full, big and fluffy they still smashed down to a proper size when I laid my head down. First night in a long time I did wake up with a headache. I was extremely happy with these pillows.
Now cooling ….I didn’t really find a difference from a reg pillow but, that again just could be me. Hope this helps
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Reviewed in the United States on May 7, 2026
★★★★★ 5
Heavenly Sleep Upgrade – DIORIS Pillows Are a Dream Come True!
I’ve finally found the perfect pillows! The DIORIS Queen Pillows Set of 2 has completely transformed my sleep experience. From the moment I laid my head down, I felt like I was at a luxury hotel — but right in my own bedroom!
These pillows strike the ideal balance between softness and support. The premium down alternative filling cradles the head and neck gently, yet offers just the right amount of firmness. Whether I sleep on my side, back, or stomach, these pillows adapt perfectly and keep me comfortable all night long.
The slow-rebound, breathable fiberfill is a game changer. It stays cool, supports proper posture, and even helps reduce neck pain — something I’ve struggled with for years. I wake up feeling refreshed and ache-free, which is priceless.
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Reviewed in the United States on August 13, 2025
★★★★★ 5
Wonderful
Size: Cooling Queen, Size: Cooling Queen
I absolutely love these pillows From the very first night, I could feel the difference. They are incredibly comfortable, soft, and still provide great support for my neck and head. Plus, the modern design makes my bed look so much cleaner and more elegant. The fabric feels high quality, and the pillows keep their shape really well. I genuinely feel like I’m sleeping better since I started using them. Great value for the price, and I would definitely buy them again. Highly recommend
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Reviewed in the United States on May 27, 2026