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Description
Human STING 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 Stimulator of Interferon Genes (STING) capture antibody. 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 Stimulator of Interferon Genes (STING) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Stimulator of interferon genes ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Transmembrane protein 173 (TMEM173), also known as stimulator of interferon genes (STING) or MPYS, is a protein encoded by the STING1 gene. It plays a crucial role in innate immunity. When cells are infected by intracellular pathogens such as viruses, mycobacteria, and intracellular parasites, it induces the production of type I interferons. The type I interferons mediated by TMEM173 protect infected and neighboring cells from local infection by binding to the same cell that secretes it (autocrine signaling) and nearby cells (paracrine signaling). | |||||||||||||||||||||||||||||||||
| 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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4.4 ★★★★★
Based on 19 reviews
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Product Reviews
★★★★★ 5
This book will help you navigate the teen years and stay in touch with your kids as they grow into young adults
I first read "The Five Love Languages" for adults and it opened my eyes to a new world. I discovered how the way I was showing love to my husband really wasn't communicating love to him very effectively because MY love language was different than HIS. Then we had a family and my three boys became teenagers and surprise surprise it was a challenge! A friend suggested this book and I immediately was drawn in by it because the concept is the same. How I was trying to communicate love to each one of my three teen and tween boys was not being received how I meant it because their love languages were different than mine. Aargh! That meant I actually had to spend the time learning their love languages and trying to understand them. This was a new idea for me, especially since by then I was a single mom and all I could think about was surviving day to day, not really trying to figure out how each teen boy thought, processed and communicated. Raising young kids I had found to be pretty easy. Teens were another ballgame altogether! Fortunately this book drew the idea of the teenage love languages together for me very easily, and at least helped put me on the right track. I started talking to each of my boys about how they thought and communicated, what they liked and didn't, and we even did the teen love language test together! They thought it was stupid, but we laughed about it and it was helpful! The five languages are, of course, the same as for adults: words of affirmation, physical touch, gifts, acts of service and quality time. I learnt that one of my sons absolutely did not like to be touched but valued when I did things for him (acts of service). I learnt that another really valued quality time even if we didn't talk much. A third needed words of affirmation and negative words tore him apart. Amazing - my languages are words of affirmation and gifts, so there was a lot of miscommunication going on! We started to establish some family rules, and, amazingly, things began to change.
I wrote in another review about teenage boundaries that my oldest two teenagers just left home this year. One started college last Fall and the other graduated HS this spring and started a full-time job for a year before he goes to college. They have grown into amazing young adults, and every time I see them they seem to be maturing more. Not everything I did was wrong - something was right, and we're STILL communicating, which is incredible and definitely thanks in part to this book! My youngest son, almost 16, is in full-blown teen years and I'm about to read this book again. Plus I now have two teenage stepdaughters and I've seen that there's a chapter here for blended families which is going to be absolutely invaluable.
In short, this is an incredible book and well worth the investment of time and money. Learning to communicate with your teens reaps rewards not only during the younger teenage years, but on through their young adulthood and I'm sure well into the future. In my opinion, it's fairly easy to raise young children, but teenagers are the greatest challenge ever presented to the modern parent given everything we and they face today. This book will help you navigate those years and help you stay in touch with the greatest gift you have been given - your kids!
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Reviewed in the United States on April 6, 2016
★★★★★ 5
Must read for parents of teenagers to learn their kid's personal love language. Teens especially need family love and support!
This is a great book to read if you have a teenager or pre-teen since those hormones and tendencies are already there. My teenager had to do a class that included this book and was able to articulate their love languages. I read this book and understand better now what translates to love for a child may not be the same thing as a parent, especially priority-wise. Example - Affection may be love language to the parent, but the teenager may prefer positive encouragement or affirmations versus hugs and pats. I think it's a great way to bridge the gap and love on your teenagers. Teenagers can be difficult, as I know personally, but they especially need to be loved on by their families in the ways they will accept it (read the book) since they are going through tremendous growth, hormone bursts, self-esteem and identity issues and more to get them through it.
This book is an easy read and well worth it if you are looking to better communicate and bond with your teenager, making sure their needs are met. You can also share your needs with them as well so they understand and respect that, i.e. if a parent has a tendency to want to hug and the teen isn't overly fond of it, there can be a compromise between them. :)
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Reviewed in the United States on April 21, 2016
★★★★★ 5
A must read if you have a teenager!
Format: Paperback
I bought this book after reading the 5 love languages as I have a son that is almost a teenager. This is a must read for any parent with an up and coming teenager as it really does help you better connect with them and understand their love language to ensure their love tank is full. When their love tank is full they really do behave better and are a lot more respectful!
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Reviewed in the United States on September 8, 2024
★★★★★ 5
The 5 love languages themselves are simple enough to "get
As an educator of 25 years plus experience, I always go back to reading and re-reading this timeless book by Dr. Chapman. What a blessing it is for the parents of teenagers and those who work with teens. The 5 love languages themselves are simple enough to "get," but putting them into practice can be a bit difficult for parents. Have no fear: there are plenty of examples and case studies in the book that will "spark" ideas or plans in the reader's mind. The last few chapters on how to deal with parental anger and different family structures are also very helpful. My only one regret: I wish someone had recommended this book to me when my kids were 9 or 10 years old. Get this book. It can be a game changer.
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Reviewed in the United States on May 2, 2016
★★★★★ 4
helpful and timely
I love the tone of this book. The author comes across well, with the whole goal being to get to know our teens at the heart level.
I found myself wanting to see my kids in this same light, as wonderful people God positioned in my life for me to know and interact with. Just that part would have made this book worthwhile. It really did help to change my heart toward them.
There were some areas where I felt entitled to respect from them, but those thoughts were dashed as I realized I owe them only love. The author also addressed every question that came up, such as, "Isn't that indulgent?" "What about discipline?" "If I just love them, won't they walk all over me?" Those questions proved I just didn't understand the meaning of the word Love. These questions were particularly answered in the chapters about acts of service and gifts, two love languages that can easily be misused by both parents and teenagers. I learned some things I'd never thought of, such as making a ceremony out of gift giving, in order for it to be accepted as a gift rather than just something a son or daughter is entitled to. About acts of service, I appreciated the advice to serve, all the while explaining what we are doing, so that they will never misunderstand and think they are entitled to acts of service, and also that they will not miss what we are doing is expressing love.
Something else that was new to me, and I hadn't counted on, was the moodiness of teenagers. I didn't know that was common and that it will be outgrown. I know, I should have known, I was obviously one at one time, but I missed the obvious, and had some aha moments personally, when I read this book. I hadn't known that there is no need to be offended or intimidated by these moody spells, but to lovingly address the person just as if they were not.
There was a whole chapter devoted to finding out our teens' love languages, and even how they might have changed dialects by growing up.
The book is up to date, acknowledging that there are cell phones and electronics, and how this affects today's teens so much more than the previous generation.
The reason I don't give it five stars is because I thought the author could have used more examples of interests kids could have. So many of the examples and suggestions involved either peer relationships, homework, or sports, all areas where I thought kids would be displaying false personalities. I would have liked to read more examples about family activities in the home, where everyone tends to be more like themselves, and where the reality of their life will lie as soon as they graduate from school. It could also be that we are farming, and our lifestyle is very home centered instead of business trip/9 -5 hours oriented, and I just couldn't relate to those examples. Anyway, that small concern didn't take much away from a great book, There is so much wise advice here, that we could easily fill in our own examples from our life. This is a book I'm glad to have in my library and one that I will read again.
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Reviewed in the United States on June 25, 2013