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Description
Mouse PKCd ELISA KitProduct Specification Usage Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High precision pipette and gun tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37 constant temperature box 4. Distilled water or deionized water Sample processing and requirements: 1. Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4 overnight, then centrifuge at 1000g for 20
Product Specification
| Usage |
Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High-precision pipette and gun tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37℃ constant temperature box 4. Distilled water or deionized water Sample processing and requirements: 1. Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4℃ overnight, then centrifuge at 1000×g for 20 minutes, and take the supernatant, or store the supernatant at -20℃ or -80℃, but avoid repeated freezing and thawing. 2. Plasma: Collect the specimen using EDTA or heparin as an anticoagulant. Centrifuge the specimen at 1000 × g for 15 minutes at 2-8°C within 30 minutes of collection. The supernatant can be assayed or stored at -20°C or -80°C, but avoid repeated freezing and thawing. 3. 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 the tissue and mince it. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1 g of tissue sample to 9 mL 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 the supernatant can be assayed. 4. 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 100 ng/mL). Then dilute to the following concentrations: 100 ng/mL, 50 ng/mL, 25 ng/mL, 12.5 ng/mL, 6.25 ng/mL, 3.125 ng/mL, 1.5625 ng/mL, and 0 ng/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each tube. Pipette 500uL of the 100ng/mL standard working solution into the first EP tube and mix thoroughly to make a 50ng/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 capture antibody against Protein Kinase C Delta type (PKCd). 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 Protein Kinase C Delta type (PKCd) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Protein Kinase C Delta type ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Protein kinase C delta (PKCδ), also known as PRCKD or NPKC-delta, is an enzyme encoded by the PRKCD gene. The protein encoded by this gene is a member of the protein kinase C (PKC) family. PKCs are a family of serine- and threonine-specific protein kinases that are activated by the second messenger diacylglycerol. PKC family members phosphorylate a variety of protein targets and are known to participate in numerous cellular signaling pathways. PKC family members also serve as primary receptors for phorbol esters, a class of tumor promoters. | |||||||||||||||||||||||||||||||||
| 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 | 1.56-100 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenates and other biological fluids |
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4.6 ★★★★★
Based on 9 reviews
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Product Reviews
★★★★★ 5
Skeptical at first, but after first use (so far) it’s a solid product
Style: 22", Color: Grey, Size: Queen
I initially bought this to stay at my MIL’s senior community (to help her get settled), so I didn’t want to spend a lot of money on it and I planned to leave it for other relatives that might want to visit. I expected it to be cheap-ish quality and probably not last through my two week stay.
First off, set up was a breeze. I laid it out and started inflation and it took about 4-5 minutes to fully inflate. The motor made noise, but it’s to be expected from something like this—I can say that it wasn’t as noisy as our Costco-bought mattress.
Second, it’s big—but that’s not a bad thing as much as I thought anyway after I saw it fully inflated. Turns out, for me, it’s a good thing. Our Costco mattress sits low to the floor and as it deflates a bit during the night, it’s tough to get up. This one (the 22” model and the largest one at this writing) is wonderful as I can sit on the edge of it and stand up easily. Your experience may be different, but for older folks it makes it really easy to get out of bed in the morning.
How does it sleep? Well, since I was visiting by myself and my significant other wasn’t with me, I can only speak for a single sleeper. It was really comfortable! It did take a bit of tweaking though as I thought I would like it more firm as I like to side sleep. The first couple of nights were a bit rough as I tossed and turned a bit in my sleep—I found that I couldn’t get comfortable on my side. For the reviews that say that it deflates after sleeping on it, I can say that it did seem to lose some air after the first two nights. The manufacturer does state that it does “stretch” and not deflate. As I wanted to test that statement, I didn’t change the firmness of it by re-inflating it to where I had it. The third and fourth nights were better sleep and I didn’t feel like the mattress deflated at all, so I found the stretch statement to be valid—it wasn’t losing air. After that night, I re-inflated it to where I thought I’d like it. The next night was the same tossing and turning that I had those first two nights, so I concluded that I needed to make it less firm and I actually DEflated it a bit and for the rest of the stay it was perfect! I didn’t have to touch it and I even unplugged it.
When I was getting ready to leave, I started the deflation process by turning the dial in the opposite direction. It took longer to deflate, so I walked away for a bit—I’d guess about 7-8 minutes. When I came back the mattress was flat as a pancake and ready to fold. This is the only hassle of the process as folding it to get it into the storage bag and back into the box (for storage). I do like the tote that they provide as the handles make it easy to lug about.
So, the first use was a success. If repeated uses reveal issues, I’ll report back. I will say that I plan to buy another one to replace our beloved Costco air mattress—she’s showing her age and the height of this one as well as the cost make it a no-brainer to get it as a second. We’ll use this one until it dies (if it even does) and then fall back on our old one if needed. I gave an overall 5 stars for the product with 4 stars for sleep quality and comfort only because your idea of these two might be a bit of a learning experience until you find your inflation sweet spot—so it’s not just an “inflate fully and use” type of scenario.
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Reviewed in the United States on May 2, 2024
★★★★★ 4
Comfortable, high-rise air mattress with built-in pump
Style: 22", Color: Grey, Size: Queen
The Intex Dura-Beam 22-inch Queen air mattress is one of the better inflatable beds I've used. The built-in pump is very convenient — no need for an external pump or battery-powered inflator. It inflates to full firmness in a few minutes and deflates just as quickly. The 22-inch height makes it easy to get in and out of, similar to a regular bed height. The plush comfort top adds a softer surface feel. The 600 lb weight capacity is reassuring for heavier users or couples. I've had guests sleep on this for multiple nights and they reported sleeping well. It held air overnight without significant deflation. My only note is to store it carefully to avoid punctures from sharp objects. Overall a great value for guests or camping.
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Reviewed in the United States on April 15, 2026
★★★★★ 5
Very good
Style: 18", Color: Grey, Size: Queen
I recently purchased this inflatable mattress from Amazon, and I’m honestly very impressed with the quality and comfort. It inflated quickly and was very easy to set up. The material feels durable and strong, and the mattress stayed fully inflated throughout the entire night without losing a
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Reviewed in the United States on May 19, 2026
★★★★★ 5
This is a very good air mattress
Style: 13”, Color: Grey, Size: Twin
I love this mattress so much I work overnights and I needed something to sleep on so I bought this mattress and it was the best mattress ever it’s so comfortable I purchased it Dec 24th2025 and it’s now starting to deflate I can’t find any holes in it but I still use it. I’m not gonna get rid of it because I can still sleep on it I just need to inflate it two times throughout the night it doesn’t bother me because it’s so comfortable. I’m not gonna lie I did purchase another one. But I’m 125 lb but the grands can sleep on it when they stay over so I will be keeping it. This is a really good air mattress well worth your money.
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Reviewed in the United States on April 25, 2026
★★★★★ 5
Decent air mattress if you take care of it
Style: 22", Color: Grey, Size: Queen
I’ve been buying this air mattress, and its previous model, for the last four years. I used to go camping and attend festivals about six times a year, and I wasn’t particularly nice to my air beds. Most of the time I didn’t fold them back up neatly or put them in the storage bag. I’d just deflate them, throw them in the car, and head to the next event. Even with that treatment, they held up surprisingly well.
I was nervous about buying the new version with the redesigned inflate/deflate module, but on a whim I decided to give it a try.
I used it during a five-night camping trip near Tygh Valley, Oregon at the end of May 2026, where temperatures ranged from about 85°F during the day to around 40°F at night.
Did it lose some air over the weekend? Yes. Was it enough to bother me? No. In fact, over the entire five-night trip, I only had to add air once. Considering the temperature swings, I was impressed with how well it held up.
The new model does seem to take a little longer to deflate than the older versions, but that’s a minor complaint. Overall, I’m very happy with it.
I’ve had good luck with these mattresses over the years, and this new version seems just as solid as the previous model.
I would absolutely recommend this mattress and will continue buying this model when it’s time to replace mine.
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Reviewed in the United States on May 31, 2026