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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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4.5 ★★★★★
Based on 17 reviews
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Product Reviews
★★★★★ 4
Okay serum, not as good as the cream though
I am obsessed with the cream version of this so I thought I’d give this a try. The dropper top allows for easy application. The formula is light and non greasy and absorbs easily. No harsh fragrances. I would use it when I had eczema flares on my face and feel like it didn’t work near as well in helping with skin irritation as the cream does. As of now I don’t use it as part of my regular skin care. Do I think it’s a bad product? No, but it didn’t really work wonders for me. Not sure I would purchase again.
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Reviewed in the United States on May 21, 2026
★★★★★ 5
Helps with redness and softens skin
I have sensitive, mild rosacea skin. Had a facial recently and the aesthetician suggested I get a serum to help with moisturizer as I am almost 40. I’ve tried a lot of serums and either they are sticky and not really hydrating or break my skin out in a rash. This serum is amazing. Helps calm the redness in my cheeks and my skin is moisturized and soft all day. It feels a little sticky at first but it goes away. I apply in this order OSEA sea mineral mist toner to prep the skin, this serum and top off with Chanel La Solution 10 for sensitive skin.
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Reviewed in the United States on December 27, 2025
★★★★★ 5
Sensitive skin
Such a good product. I use after micro needling when skin is sensitive. Also a great product to use as a barrier serum when starting tretinoin cream.
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Reviewed in the United States on April 24, 2026
★★★★★ 5
Saved my skin during an eczema flare up
This saved my skin!
I had an eczema flare on my face, and my usual products weren’t working. A friend recommended the Avene brand and within 3 days of using this rescue barrier serum, my flare was almost completely gone! This saved my face!
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Reviewed in the United States on April 11, 2026
★★★★★ 1
PLEASE REVIEW — FORMULA CHANGED!!!
I wish I would’ve posted a review PRIOR to the formula changing but just know that I LOVED this serum. I’ve struggled with hormonal acne (PCOS) and eczema for as long as I can remember. Finally, last summer my dermatologist put me on (3) different RX including Spironolactone and Accutane. I was so tired so I stopped everything except for CeraVe cleanser, retinol (Tretinoin) and CeraVe healing ointment. My made up skin routine was working until I used too much Tretinoin and burned my face. I had read great reviews about this product so I thought, I might as well try it — and IT WORKED! I’ve been using this product for almost a year and I loved it. However, 3 weeks ago when I ordered it, I noticed the bottle had a green tint. I could tell after one use that the feel of the serum was different, more watery, not as thick as it used to be. The smell was also different. Still, I kept using it for nearly 2 weeks and I experienced 2 breakouts! Sure enough, I stopped the product and within a few days, my skin was back to normal. I am very upset because as someone who has extremely sensitive skin, this worked but unfortunately, the new formula isn’t for me.
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Reviewed in the United States on May 14, 2026