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Description
Human PRKG1 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 ultrasonically disrupted or repeatedly frozen and thawed. Finally, the homogenate is centrifuged at 5000×g for 5-10 minutes and the supernatant is collected 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 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 utilizes a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA). Sample, standard, biotinylated detection antibody, and HRP conjugate are sequentially added to microwells pre-coated with a capture antibody against Protein Kinase, cGMP Dependent Type I (PRKG1). After incubation and washing, the sample is developed using the substrate TMB. TMB converts to blue under the catalysis of peroxidase (HRP) and to yellow under the action of acid. The intensity of the color is positively correlated with the amount of Protein Kinase, cGMP Dependent Type I (PRKG1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Protein Kinase, cGMP Dependent Type I ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | cGMP-dependent protein kinase 1 (PRKG1) is an enzyme encoded by the PRKG1 gene. It is a protein-coding gene. Diseases associated with PRKG1 include aortic aneurysm, familial thoracic septal 8, and familial thoracic aortic aneurysm and aortic dissection. Pathways involved include apoptosis and survival, anti-apoptotic effects of nuclear ESR1 and ESR2, and cytoskeletal signaling. This gene is associated with autosomal dominant thoracic aortic aneurysm and dissection (TAAD). | |||||||||||||||||||||||||||||||||
| 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 and other biological fluids |
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4.1 ★★★★★
Based on 23 reviews
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Product Reviews
★★★★★ 3
below average value but decent quality on this 95.4% Polyester, 4.6% Spandex dress shirt
Special Size: Black Buttons, Size: Large, Color: Sky Blue, Special Size: Black Buttons, Size: Large, Color: Sky Blue
At $20 when I picked this up, it was overpriced for what it was. I'm held accountable to the value assigned to me at the time I select to review it, so I review the value based on what was assigned not on what it later becomes if they discount it. As I write this I can see it is in a value range it should have been for me, but I'm still reviewing it at the $20. That's overpriced on a 95.4% Polyester, 4.6% Spandex shirt (i.e., non-cotton) and so the value here is a 1/5 or 2/5, say 1.5/5 for it. In terms of quality, this is a poly dress shirt. Still, I'd call this a 4/5 for what it is.
The standard is cotton, and in my experience, as someone in ties and collared shirts all day, I would note that I do prefer a 95 cotton/5 spandex blend, with the challenge that its not the most wrinkle free material. This blend, 95 poly/5 spandex, is a very wrinkle free and smooth feeling material. Poly has better durability than cotton despite feeling thinner, because it won't fray as much. That said, even though it's lightweight, poly will make you hotter because it doesn't breathe like cotton. It feels light, so you think it is going to be cooler, but in practice actually isn't really if it's a hot day. It can make you sweat more.
That said, many people like poly blends for the ease of maintenance. I've given up dry cleaning, just too much, and I do like to avoid an iron, so I tested this out because of it. For me this fit well. I ordered a large (typically I prefer to wear a 16.5/34/35, and I'm 5'11, 185) and this fit well. It's not a slim fit (tapered bottom) for those who might complain about extra material. But it fit well. I put it through both the wash and dryer. It did not shrink (poly usually doesn't) to be out of size, but was fine, and of course came out without wrinkles. The collars have no stays, these poly shirts hardly ever do, but I have found they mostly remain flat, since the material doesn't tend to wrinkle, and thus my points stay in place just fine.
At a 1.5 on value and a 4 on quality, this is a 2.75/5 average for me. Again, value plays a big part for me. So, full disclosure a better value would have raised my rating on this. But hopefully, based on what I covered above, you have enough to figure out if this shirt is worth it to you or not.
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Reviewed in the United States on March 2, 2026
★★★★★ 5
Thev quality and size
Special Size: Black Buttons, Size: Large, Color: Grey
It's good fit
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Reviewed in the United States on March 21, 2026
★★★★★ 5
Nice dress shirt
Special Size: Black Buttons, Size: Large, Color: Grey
This shirt is a super soft and an awesome style, which I really love. The material is a soft cotton and that doesn’t irritate my skin. It is breathable, but also keeps you relatively warm. It’s very nice looking on and would be okay for everyday wear. It has been used many times going out with jeans or shorts out. I wash and dry it regularly in a machine and still looks just as new as the out of box condition. There is no pilling, fabric change, or shrinking. The style is flattering, cut well, and runs true to size. Other shirts owned feel similar to this one, which meets expectations and is a nice shirt for the price. I do wish it had a pocket though. Ultimately would purchase again.
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Reviewed in the United States on April 7, 2026
★★★★★ 5
Sharp-looking business shirt at a great price
Special Size: Black Buttons, Size: X-Large, Color: Rhine Blue
This long sleeve dark blue shirt is a great choice for a business casual look. The "Rhine Blue" color really stands out — it’s very bold without being over the top, and it gives the shirt a sharp, polished appearance.
The stitching is very well done, with clean lines and no loose threads, which adds to the overall quality. It fits nicely and maintains a crisp look throughout the day.
The material is mostly polyester with some spandex, so it feels smooth, lightweight, and is actually breathable. It stays comfortable during long wear and doesn’t feel heavy or restrictive.
Overall, it’s a sharp, well-made shirt that delivers both style and comfort at a great level. It was a good deal at the original price of about $20, but at the new price of about $15, it's an excellent deal. I may get another in a different color.
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Reviewed in the United States on March 29, 2026
★★★★★ 5
Very comfortable and looks great!
Size: X-Large, Color: Navy Blue, Size: X-Large, Color: Navy Blue
This shirt is surprisingly really nice for the price. Really nice even regardless of the price. It fits and looks great on my hubby. We liked it so much that I bought him another one in a different color. Will probably even buy more. The large ran a little small, so we got him an extra large and it was great.
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Reviewed in the United States on February 6, 2026
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