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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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4.8 ★★★★★
Based on 1445 reviews
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Product Reviews
★★★★★ 5
Best pillows we’ve ever had!
Color: White, Size: King(Pack of 2)
These are so insanely comfortable! They have the perfect amount of softness and support. Not too flat, not too full, just right. You will sink into it like a big marshmallow!
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Reviewed in the United States on May 22, 2026
★★★★★ 5
Comfortable pillows 🤩
Color: White, Size: Queen(Pack of 2), Color: White, Size: Queen(Pack of 2)
I bought the Serta Hotel Collection Bed Pillows (Queen size) and I absolutely loved them. They are super soft yet provide great support; they feel very comfortable and help make for a much better, more pleasant night's sleep. Plus, the fabric is cool and of high quality. I recommend them wholeheartedly!
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Reviewed in the United States on April 22, 2026
★★★★★ 4
Amazing pillows!
Color: White, Size: Queen(Pack of 2)
These are fantastic pillows! A little background. I'm a side sleeper and toss and turn throughout the night. I've struggled with sleep issues for literally decades - insomnia, tossing and turning, waking up multiple times a night to use the bathroom, waking up way too early and not being able to get back to sleep. Every night it's a combo of those things or all of those things. I've also had some neck pain and stiffness for a couple of months. If you've ever dealt with that you know it will affect sleep which will then affect productivity and mood. It's a terrible spiraling and compounding thing to deal with, very stressful. I figured the contour pillow I had which once was actually helping was unfortunately the culprit. So I went on the hunt for another pillow. I ordered what I thought was the old Rejuvenite pillow (those were fabulous!) and got burned on a bait and switch. The pillow ended up being way too soft and gave me worse neck pain. Figuring I'd try to salvage what was a $115 pillow, I invested in a very thin pillow to put underneath. It was too high. Getting pretty frustrated, I ordered the cheaper Utopia pillows here on Amazon. Those actually were not terrible because they did take away my neck pain, but I was still tossing and turning at night and still feeling unrested in the morning. I decided I'd give it one last shot and luckily found these when they were discounted by a few bucks. They didn't have many reviews which was actually a great sign for me because it means the company lets their product speak for itself.
I got these in several days and was thrilled to see upon opening the package that they were sealed and the directions do not require you to open the package and let them expand for 24 hours. You literally just take the edges and fluff 2-3 times and that's it! They are ready to go and they definitely do feel perfect after doing that. I was skeptical when I put them on my bed because they are soft and I worried they'd be too soft. I felt my head sink into the pillow a bit and then was surprised to feel at a certain point it just feels like the pillow is cradling your head with the right amount of support. I wouldn't describe it as firm, just soft but stable support. Really comfortable.
But the real test was how it would perform during sleep. I'm so thrilled to report that I had an amazing night of sleep! No neck pain, no stiffness, zero tossing and turning, no insomnia, no waking up during the middle of the night, and no waking up early. I put my head down, fell asleep, and slept like a log for about 7.5 hours straight, which is literally unheard of in my world. I'm very lucky if I get 5 solid hours of sleep. I woke up and popped out of bed. My mood was great (still is later in the day) and I was not tired in the afternoon when I normally feel the fatigue kicking in. This is the first day for as long as I can remember that I do not feel that nagging tiredness behind my eyes and in my body that always seems to be a steady background actor in my life. I feel great and full of energy and I was able to tackle a bunch of tasks on my someday list. This is so promising I didn't want to wait to share. If you are on the fence and are dealing with similar issues to mine, I'd encourage you to just try these.
What's also amazing is these come in a set. I'm only using one so I basically have another one to use if the first one ever fails. This is the best investment for sleep I've ever made!
Update:
Well, after a trial period I'm removing a star because my dreamy sleep was short lived. I started to wake up more frequently during the night with these and gave up midway through the night one night. I'm still using these for back support while I sit up or hang out in bed because they are still super comfortable that way and oddly these work well for naps. Hopefully, you have better luck than me!
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Reviewed in the United States on February 21, 2026
★★★★★ 5
Absolutely love this pillow!
Color: White, Size: Queen(Pack of 2), Color: White, Size: Queen(Pack of 2)
The pillow was very well packaged and extremely easy to open. As soon as I took it out of the package, it immediately expanded and became fluffy within seconds. I was honestly surprised at how quickly it regained its full shape.
The pillow feels very soft and comfortable, but it still provides great medium-firm support. It has excellent resilience and bounces back quickly after being pressed, so it never feels flat. I’ve been using it both for sleeping and for supporting my lower back when sitting, and it works great in both situations！
I also really like the embroidered logo, which looks very delicate and well made, it adds a nice premium touch to the pillow. It would also make a great gift for family or friends.
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Reviewed in the United States on March 6, 2026
★★★★★ 5
Heavenly soft and comfortable.
Color: White, Size: King(Pack of 2)
This is a great pillow. I bought 2 king size for my daughter. They are so soft but have some heft to them. They don't squish flat. They plump up so nice. She said like having a cloud for a pillow. I love Serta. They have always been good. Buy it and sleep like you're in heaven.
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Reviewed in the United States on June 2, 2026