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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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4.5 ★★★★★
Based on 15 reviews
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Product Reviews
★★★★★ 5
A Classic
Format: Mass Market Paperback
A classic that deserves to be re-read to our children and grandchildren
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on March 22, 2026
★★★★★ 4
A challenging read
Every now and then I challenge myself with a classic. This one was definitely that - a challenge. Allow me to explain.
First, this is the story of King Arthur. All the characters are present: Gweniviere, Lancelot, Merlin, the Knights of the Round Table, the sword in the stone, Mordred, and all the ones I’d ever heard of in retellings-of the story. However, many were not as good, bad, beautiful, handsome, or courageous and pure of heart as I’d been led to imagine. These characters were messy and deep with honor and dishonor battling within them. But there was a quiet strength and desire to do good about them. So, this book challenged my preconceived images of these characters.
Second, this book is long. It is divided into four parts. The first two parts were more familiar (maybe) and were a little easier to read. They told of Arthur’s early education under Merlin, pulling the sword from the stone, and establishing the Round Table - the glory days. Part three tackles the love triangle between Arthur, a Gwen, and Lancelot; the quest for the Holy Grail; and the rise of Mordred. Part four got a little tedious describing the fall of Arthur’s reign, the destructive force of lies and secrets, and the power of evil. This section challenged me with some very deep theological and philosophical musings amid the battles among friends and family.
Finally, this book is written using a Middle English wordings and geographic dialect. I had to look up many words (not all definitions could be found) to really understand some things, but was generally able to follow the story. The author also frequently talked directly to the reader which took some getting used to. The writing style definitely challenged my vocabulary and ability to follow transitions from author speaking to characters.
One last thing, the ending was left somewhat open to interpretation and imagination. I choose to believe the happiest of the possible outcomes. I’m glad I read this book, but i think I’ll go read something a little easier next!
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Reviewed in the United States on March 22, 2020
★★★★★ 5
Freshest breathe
I recently purchased a stainless steel tongue cleaner, and I am pleased to report that it is a great product. The cleaner is made of high-quality stainless steel, which is extremely durable and will not corrode over time. It is also easy to use and clean, as the cleaning part is made of a smooth and rounded end that fits comfortably in my mouth.
The cleaner is also very effective at removing bacteria and food particles from my tongue. After using the cleaner for a few days, I noticed that my breath had improved significantly and that my taste buds were more sensitive. I was also surprised to find that the cleaner was gentle enough to use on an everyday basis without causing any irritation.
Overall, I am very happy with my purchase and would highly recommend it to anyone looking for an effective and affordable tongue cleaner. Not only is the stainless steel cleaner effective and easy to use, but it also lasts longer than other tongue cleaners on the market. I would definitely recommend this product to anyone looking for an effective and reliable tongue cleaner.
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Reviewed in the United States on May 10, 2023
★★★★★ 5
Nice product
This is a great tongue scraper. Its metal and is a two pack. Definitely worth the money. I di recommend this product.
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Reviewed in the United States on May 27, 2026
★★★★★ 5
Exactly what I needed
These are solid and work really well. They’ve held up perfectly and as long as I don’t lose them I can’t imagine them wearing out for a really long time.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 25, 2026
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