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Description
Human PCSK5 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: Adherent cells should be gently washed with pre-chilled PBS, then trypsinized and harvested by centrifugation at 1000×g for 5 minutes. Suspension cells can be harvested directly by centrifugation. Collected cells should be washed three times with pre-chilled PBS and resuspended 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. 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 capture antibody against proprotein convertase subtilisin/kexin type 5 (PCSK5). After incubation and washing, the sample 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 amount of proprotein convertase subtilisin/kexin type 5 (PCSK5) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Proprotein Convertase Subtilisin/Kexin Type 5 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Proprotein convertase subtilisin/kexin type 5 (PCSK5), also known as PCSK5 or simply proprotein convertase 5 (PC5), is an enzyme encoded by the PCSK5 gene. The protein encoded by this gene belongs to the subtilisin-like proprotein convertase family. Members of this family are proprotein convertases that process latent precursor proteins into their biologically active products. This encoded protein mediates post-translational proteolytic processing of several integrin α subunits. It is thought to process pronephrin, cytokine type 1 matrix metalloproteinase, and HIV-1 glycoprotein gp160. Mutations in this gene are associated with Currarino syndrome-like malformations. | |||||||||||||||||||||||||||||||||
| 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, and other biological fluids |
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4.5 ★★★★★
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Product Reviews
★★★★★ 5
Life changing book
Format: Paperback
Amazing life changing book! I’ve grown so much from reading this book than I have in years. It has opened my eyes to learn more about myself and others - that others have recognized that I’ve changed. I hope others discover the same impact as it has on me!
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Reviewed in the United States on April 4, 2026
★★★★★ 5
Self-Care’s Always in Season
Format: Paperback, Format: Paperback
Self Care for Adult Children of EI Parents is a great book to teach the lessons in life you maybe never got the chance to learn. How can you know how to handle or deal with things you were never taught properly or modeled?
This is your start right here! Identify places you need to grow. Self-care comes in several forms and in this book you can explore where you may know you need help or start on page one. Often times caring for others is the way your brain is wired based on experience. Throughout this book you will find tools to get you on track putting yourself care first so you are able to show up for those who deserve if and conserve the energy on those who don’t.
It’s also a great addition to the set by Lindsey Gibson for Emotionally Immature “Parents/People”.
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Reviewed in the United States on January 15, 2026
★★★★★ 5
Life Skills You Were Never Taught
Format: Paperback
I have never felt so seen and heard by Dr. Gibson. This book has truly been life altering for me. The lessons in this book have helped me become a confident leader and a more stable, happy, and kind person. I understand myself so much better than before, and now see how my upbringing has affected me into adulthood.
The valuable lessons in this amazing book are easy to understand and implement in everyday life. Lessons are categorized and laid out in a very easy to read and retain way.
I also have the audiobook and love to follow along and highlight passages. The hearing of the kind words in this book is so healing for me, and I highly recommend it. I feel motivated by this material each time I see it.
Thank you so much, Dr. Gibson, for you dedication to this topic. It has sincerely changed my life for the better.
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Reviewed in the United States on June 30, 2023
★★★★★ 4
Therapist recommended
Format: Paperback
My therapist recommended this book, and I can already see why. It’s incredibly validating and insightful. It really helped me understand so many of my feelings and behaviors in a way I hadn't before. The exercises and practical advice are so helpful and feel very grounded. It's a must-read if you're working through this kind of stuff. I’m so grateful my therapist told me about it.
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Reviewed in the United States on August 28, 2025
★★★★★ 5
Great book for direction on how to help yourself!
Format: Kindle
This book, written for (as the title suggests) adult children of emotionally immature (EI) parents, provide wonderful insights into the psychology of EI people. Regardless of the emotional level of your parents, these books are a must-read for everyone. Why? Gibson kindly and directly delivers clear insights into EI behavior, which applies to any adult in your life.
Self-Care for Adult Children of Emotionally Immature Parents: Honor Your Emotions, Nurture Your Self, and Live with Confidence, deals with caring for yourself in ongoing relationships with EI people. These techniques are grouped by where and when you may encounter EI people to use the book as a reference guide. If you are close to someone who exhibits EI behaviors, you can flip to that section. If you work with EI people, there's a section for coworkers. This book focuses on how you nurture yourself and your identity once you've worked on healing from childhood trauma.
Gibson does a superb job of sharing direct context about EI people. She pulls no punches about EI people's damage but gives space to encourage growth and break the multi-generational trauma.
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Reviewed in the United States on July 21, 2023