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Description
Mouse TAF12 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 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 transcription initiation factor TFIID subunit 12 (TAF12). After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by peroxidase (HRP) catalysis and to yellow by acid. The intensity of the color is positively correlated with the level of transcription initiation factor TFIID subunit 12 (TAF12) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Transcription initiation factor TFIID subunit 12 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | TATA box binding protein-associated factor 12 (TAF12), also known as transcription initiation factor TFIID subunit 12, is a protein encoded by the TAF12 gene. Control of transcription by RNA polymerase II involves the basal transcription machinery, a collection of proteins. These proteins assemble into complexes with RNA polymerase II, which are regulated by transactivator proteins that bind to cis-regulatory elements located near the transcription start site. Some regulators interact directly with the basal complex, while others may act as bridge proteins, connecting the transactivator to the basal transcription factor. Some of these associated factors are weakly attached, while others are tightly associated with TBP within the TFIID complex. The latter group includes TAF proteins. Different TAFs are predicted to mediate the functions of different transcriptional activators of various gene promoters and RNA polymerases. They interact directly with TBP as well as TAF2I. | |||||||||||||||||||||||||||||||||
| 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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4.9 ★★★★★
Based on 16 reviews
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Product Reviews
★★★★★ 5
Great book!
Format: Paperback
Such a great book! I wish I would have had this when my kids were little, but it's also helpful for my grandkids. It's a game changer! If you are looking for a book to raise good kids, this is the one!
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Reviewed in the United States on March 11, 2026
★★★★★ 5
Wonderful book
Format: Paperback
Great book and wonderful advice for struggling parents
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Reviewed in the United States on May 14, 2026
★★★★★ 5
Please read, especially if you read the bad reviews.
Format: Paperback
I love this book... I bought it over 15 years ago... And after reading the reviews (especially the bad ones), I had to comment. Most of the comments have to do with spanking and it is obvious that if you are against it, you will have problems with it, (even though there is a TON more advice on all other aspects of parenting than just spanking). The reason for this review is because when I first read this book, I was against spanking too, but because of this book I have a new understanding of it, and have only used it once on each of my two children. If you have an open heart and mind to hear what is being taught in this book, instead of a "heels dug in" attitude, you will learn a lot about what it means to be a Christian parent and have spanking as part of your discipline, even if you never have to use it. I was invited to a parenting class at a friend's church that used this book.. I was in a place of discouragement. I felt out of control when it came to knowing what was right and wrong with raising and disciplining kids. My mother never once spanked me as a child. I spanked my son (at 2) once and that was it.. I felt horrible and abusive. I vowed never to spank again. I then became a Christian when my kids were 5 and 2.... this book put into perspective God's intention of authority. We are all under some kind of authority.. and it should be respected for various reasons... This book taught me that spanking must not be violent, done in anger or out of frustration. and the first time I used SACH practice, I explained to my then 5 year old that because he broke a rule, he would be punished and his punishment would be a spanking. To go to his room and think about what he had done and wait for me. I took about 10 minutes to settle down, prayed for God' guidance and wisdom and then went to my son's room. I told him to take down his pants and lay across my lap (he still had his underwear on) and I said, "Because you broke ....... rule, you will be spanked now." and I lightly slapped his bottom, no more than than like lightly slapping a mosquito off your hand. He cried more because he was sorry for what he had done than any "pain" from the spanking. We talked about why the rule is in place, why it is important that he follows it and that mommy and daddy make rules because we are older and wiser and only wish for his safety and happiness. I never had to spanked my child again.. he is now 20. From the moment I read this book, my husband and I started developing a relationship of respect with our kids that demonstrated our love for them out of our authority and responsibility for them. Both my kids are well adjusted and have never been in ANY kind of trouble. They know they can come to us and our relationship is deep, loving and they comment often how they recognize how special our relationship is compared to their friends. We have also acknowledged that we are human and make mistakes, and we own up to our mistakes and ask our children for forgiveness. That goes a long way with kids. but establishing respect and authority with your children will allow them to grow in a safe environment. And putting God in the center of your family showing your kids that you too are under the authority and grace of God will plant a seed of a long lasting relationship with Christ, which I believe is every parent's hope and goal... Happy, healthy, successful kids who love God.
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Reviewed in the United States on November 24, 2014
★★★★★ 5
Incredibly Biblical, Amazing Insights, Best Discipline Book on the Market. NOT ABUSIVE!!
Format: Paperback
The only reason people rate this book low is because it advocates spanking. Because spanking doesn't happen to be popular in our culture, and because people have been brainwashed to believe that spanking is abusive, people think this book is awful. If this is you, or you're not a Christian, don't even bother reading this book. It will just anger you, mostly because you won't understand it.
BUT, if you are a Christian, and you want the most Biblically based instruction on RAISING your children, then this is by far the most incredible book you could ever read. This book is not about spanking. In fact, he only talks about spanking in two chapters. This book is about WHY and HOW to discipline your children based on what the Word of God says. It has been so impactful for me, that I would be willing to live my entire life over again if I could go back and have my mother raise me as this book suggests. I would be a different person today with a LOT less problems.
The way Tedd Tripp explains authority and why we discipline our children is so different than most people have been raised. It's not because it's "out there", but because the church as mostly failed in helping us understand discipline and authority. For example (from the book), we are under God's authority, and God has commanded us to bring up our children in the training and instruction of the Lord (Eph 6:4). Our children are under our authority. So when we discipline them, it's not because we're angry or we want to 'show them who's boss', but it's because we are commanded by the Lord and we must be obedient to Him. Not only is this Godly, but it teaches our children about authority and changes the nature of discipline! This is just the tip of the iceberg on dozens of amazing concepts that I personally had never thought about before in my life.
This book is very straightforward and easy to read. Tedd Tripp isn't a flowery writer and doesn't waste words. He's direct, to the point, and often doesn't even give examples (something I would have liked more of). But because it's so straightforward, it makes sense and goes fast. I plan on reading this book several times just to ingest everything it has to say.
I hope every Christian parent reads this book; our society would change if it followed the guidelines laid out in this Godly, insightful book.
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Reviewed in the United States on November 28, 2005
★★★★★ 4
An Excellent Book
Format: Paperback
There are many things in life that are easy to do poorly but are much more difficult to do with excellence. It did not take me long as a parent to discover that it would not be difficult to raise children, but that it would be exceedingly difficult to do it with excellence. In the six years since my eldest child was born I have looked often for help and advice in becoming an excellent parent. Unfortunately my wife and I have received little mentorship in this area. Thankfully, there are many books written about this topic so we have often looked to these resources to provide the wisdom and training we know we need.
Shepherding A Child's Heart by Tedd Tripp came to us highly recommended. In fact, I can't think of a book on this topic that was recommended to us more often. It is a book that deals with speaking to the very heart of your children. Realizing that too many parents react only to symptoms of underlying sin, Tripp attempts to help parents look deeper, to see that all the things a child says and does flow from the heart, for as Luke 6:45 says, "...out of the overflow of the heart the mouth speaks." If a parent can understand a child's heart and shepherd that heart, he can deal most effectively with a child's deepest needs. And through it all he seeks to keep the gospel central to a parent's calling and to a child's response.
The book is divided into two parts. In the first, Tripp lays the foundation for biblical childrearing. He shows that the heart of bad behavior is a sinful heart. He discusses a child's development, showing that a child is shaped by various influences on his life and that a parent needs to help a child have a Godward orientation. He discusses authority and suggests that, despite our culture's disgust towards authority, a parent must assert himself as being in a position of God-given authority over a child. A child must realize that parents speak not of their own authority, but of God's. He also discusses goals, methods, communication and discipline.
Where the first part of the book lays a foundation, the second part guides a parent through shepherding a child through three stages of development: infancy, childhood and teenagers. For each of these periods he suggests the training objectives and then procedures a parent should use to attain these objectives.
A section I found particularly interesting, perhaps because I have young children, was the section dealing with punishment. Tripp advocates spanking as really the only biblical method of punishment (and certainly the only one that is specifically mandated by Scripture) for correcting young children. He lays out very clear circumstances in which children should be spanked and suggests many circumstances in which parents must not spank. He makes this type of corporal punishment very deliberate and very loving. He suggests that parents must be fully in control of themselves when they spank and must not be filled with anger. Parents do not punish their children out of anger, embarrassment or retribution, but to teach children that defying authority will bring about consequences. Children must know that God demands obedience to authority and that there are consequences for defiance.
In his endorsement of this book Edward Welch wrote, "Dr. Tripp's material on parenting is clearest, most biblically framed, and most helpful that I have ever encountered. It has become the backbone of my own parenting." I agree entirely. Throughout the book Tripp focuses on Scripture and on the gospel. He focuses on human nature and on the grace of God in providing a solution to the needs of our children. I would not hesitate to recommend this book to any parent, and especially to new parents. Read it now, pray about it, and let God direct you to His ways of shepherding the hearts of your children.
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Reviewed in the United States on August 26, 2006