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Description
Rat PKA 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: Gently wash adherent cells with pre-chilled PBS, then trypsinize and collect the cells by centrifugation at 1000×g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash the collected cells three times with pre-chilled PBS and resuspend 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. Cell Culture Supernatant: Centrifuge at 1000×g for 20 minutes. Collect the supernatant for analysis, or store at -20°C or -80°C, but avoid repeated freezing and thawing. 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 10 ng/mL). Then dilute to the following concentrations: 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 ng/mL, 0.15625 ng/mL, and 0 ng/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each. Pipette 500uL of the 10ng/mL standard working solution into the first EP tube and mix thoroughly to make a 5ng/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 protein kinase A (PKA) capture antibody. After incubation and washing, the assay is developed using the substrate TMB. TMB converts to blue under the catalysis of HRP and to yellow under the action of acid. The intensity of the color is positively correlated with the protein kinase A (PKA) content in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Protein kinase A ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Protein kinase A (PKA) is a family of enzymes whose activity depends on cyclic AMP (cAMP) levels in cells. PKA is also known as cAMP-dependent protein kinase (EC 2.7.11.11). It has multiple functions in cells, including regulating glycogen, sugar, and lipid metabolism. PKA is also often referred to as cAMP-dependent protein kinase because it is traditionally believed to be activated by releasing its catalytic subunit when levels of the second messenger cyclic adenosine monophosphate (or cAMP) rise in response to various signals. | |||||||||||||||||||||||||||||||||
| 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 | 0.156-10 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates, cell lysates, cell culture supernatants, and other biological fluids |
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4.2 ★★★★★
Based on 27 reviews
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Product Reviews
★★★★★ 5
Great Value, Versatile and Efficient
I bought this USB-C Hub 6-in-1 Adapter during summer of 2025, and it has been a game-changer for expanding my tablet connectivity options. Below is my experience with it:
• The 6-in-1 design offers a variety of useful ports, including HDMI, USB-A, USB-C, and an SD card slot. It’s perfect for connecting multiple devices to my tablet or laptop at once.
• The 4K HDMI port works flawlessly when I connected it to my TV and monitor, and the video quality is crisp and clear, with no lag or issues.
• I also connected my tablet audio via the 3.5mm (1/8") port for the audio output and it sounded great with no delay in audio.
• This 100W PD feature is fantastic. I can charge my tablet or laptop while using the hub, which is incredibly convenient when I’m working on the go. It’s great to have a single device that handles both power and data transfer.
• The hub surely is compact, sleek, lightweight, portable and easy to carry around, making it ideal for travel or working remotely.
• The awesome part is that no additional drivers or software needed—it's just plug and play, and it works right away.
Overall: The Anker USB-C Hub 6-in-1 Adapter is a reliable and versatile solution for anyone needing to expand their tablet/laptop’s connectivity. The HDMI support, 100W power delivery, and compact design make it an excellent choice for both home and on-the-go use. Highly recommended for those who need a multi-functional hub for work, presentations, or entertainment! Keep in mind that using 45W to 100W chargers are best to use but do not exceed above 100W.
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Reviewed in the United States on October 25, 2025
★★★★★ 5
Anker 6-in-1 USB C Hub
I do enjoy the features of this USB-C hub. It provides the connectivity I wanted in my truck. It quickly charges any device via 85w capable USB-C pass-through charging. It offers 10Gbps USB-A ports that transfer files quickly and easily between devices. I was informed the hub functions properly with newer devices that deliver audio through the USB-C port instead of a 3.5mm aux jack. I will have to test the USB-C audio feature some day.
Pros: affordable, build quality, availability, connectivity, ease of use, portability
Cons: none
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Reviewed in the United States on June 26, 2025
★★★★★ 5
Make Sure to Use a Power Delivery Charger with this.
This hub is pretty good. If you don't use it with a PD charger though, the USB C port will not charge your phone. Even then, it will charge pretty slow. Everything else works great.
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Reviewed in the United States on July 11, 2024
★★★★★ 4
Good adaptor that does not get extremely hot, but may have wifi issues with Macbook Air M2
Received the adaptor a few days ago. I use it mainly to connect with my Macbook Air M2. It has a cool design, but not very sleek. One advantage of this product is it does not get really hot as other adaptors I have used in the past. However, there are several other issues:
(1) I have been having a very poor wifi connection once this adaptor is plugged in. I was unable to connect to the internet at all. It took me hours to figure out the adaptor is the cause. Once I unplugged it, the problem was solved. I restarted my computer and plugged the adaptor in, the problem hasn't come yet after 2 days. Fingers crossed.
(2) The audio headphone jack port is not working at all. Not a big deal breaker.
(3) I read online that the HDMI 60 Hz is actually not supported on this adaptor. Because my external monitor is not a 4k monitor, there is no way to test this feature out.
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Reviewed in the United States on March 22, 2023
★★★★★ 5
Versatile and Efficient – A Must-Have for Connectivity Needs!
I recently bought the Jet USB-C Hub 6-in-1 Adapter, and it has been a game-changer for expanding my laptop’s connectivity options. Here’s my experience with it:
• Multiple Ports: The 6-in-1 design offers a variety of useful ports, including HDMI, USB-A, USB-C, and an SD card slot. It’s perfect for connecting multiple devices to my laptop at once.
• 4K HDMI Support: The 4K HDMI port works flawlessly. I’ve connected it to my TV and monitor, and the video quality is crisp and clear, with no lag or issues.
• 100W Power Delivery: This feature is fantastic. I can charge my laptop while using the hub, which is incredibly convenient when I’m working on the go. It’s great to have a single device that handles both power and data transfer.
• Compact and Portable: The hub is sleek, lightweight, and easy to carry around, making it ideal for travel or working remotely.
• Plug and Play: No additional drivers or software needed—just plug it in, and it works right away.
Overall: The Jet USB-C Hub 6-in-1 Adapter is a reliable and versatile solution for anyone needing to expand their laptop’s connectivity. The HDMI support, 100W power delivery, and compact design make it an excellent choice for both home and on-the-go use. Highly recommended for those who need a multi-functional hub for work, presentations, or entertainment!
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Reviewed in the United States on December 12, 2024