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Description
Human HDAC11 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 histone deacetylase 11 (HDAC11) capture antibody. 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 amount of histone deacetylase 11 (HDAC11) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Histone Deacetylase 11 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Histone deacetylase 11 is a 39 kDa histone deacetylase encoded by the HDAC11 gene on chromosomes 3 and 6. It is the only Class IV HDAC, as it does not fit into either Class I or Class II because it shares little homology with the yeast enzymes Rpd3 or hda1. HDAC11 controls DNA expression by modifying the core histone octamer, packaging DNA into a compact chromatin structure and repressing gene expression. HDAC11 expression is typically found in brain and testicular tissues, and diseases associated with it include mature T-cell and NK-cell lymphomas. Pathways involved include activated PKN1 stimulating transcription of the AR (androgen receptor)-regulated genes KLK2 and KLK3 and the BRCA1 pathway. Gene Ontology annotations associated with this gene include hydrolase and histone deacetylase activities. An important homologue of this gene is HDAC4. | |||||||||||||||||||||||||||||||||
| 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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4.3 ★★★★★
Based on 24 reviews
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Product Reviews
★★★★★ 5
I thoroughly enjoyed reading this and as the Principal of a Middle ...
Format: Hardcover
I thoroughly enjoyed reading this and as the Principal of a Middle School, I found much truth in what she has written. I found myself thinking back to my days as the parent of burgeoning, wonderful and successful adults. The suggestions Ms. Lahey offers to her readers are based in experience and in conversations with her peers and others in the field of child-rearing and education. These are ideas that have worked for her, and others, and she passes along that wisdom and experience.
Interestingly, I passed this book on to my 33 year old daughter who works in the private sector with a small, but growing, tech company. She has found the ideas presented here very valuable in dealing with her co-workers, the people who report to her, and even her boss, all of which I found very interesting.
The best thing about this book is Ms. Lahey's voice. She sounds compassionate, not absolute, "this is the ONLY way you should ...". She puts out the propositions and then provides what she found as possible courses of action, I would highly suggest that any parent of young children, any teacher and certainly any building principal read this book with great gusto!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on October 22, 2015
★★★★★ 5
Extraordinarily Helpful - A total game changer
Format: Kindle
I read this book at the recommendation of my son's school counselor. It was a total eye opener and will greatly impact and change the way I raise my son going forward. This book could have easily been titled, "The way I used to parent" because it is so accurate down to so many details as it describes my parenting techniques and motivations. I have made many changes already and my son doesn't quite understand who I am anymore. I'm certain that he appreciates the changes although he's a bit disoriented currently, but I know he will be better off in the long run.
I wish there were more anecdotes or perhaps several role playing scenarios added to help offer more suggestions on how to handle more situations but the book was extremely helpful and insightful regardless. Most of the anecdotes I saw play out in our family or with other students from my son's school within hours or days of reading the book. It was amazing.
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Reviewed in the United States on November 22, 2016
★★★★★ 3
Good book for parents
Format: Hardcover
A good review for parents to keep as they struggle, but as reviewer noted a bit oversimplified.
I bought after had listened to a panel on Forum discussed screen time. As a grandparent, sitting with my wife each on our I pads, I recall eating at restaurants when you look an see so many n smart gadgets texting, tweeting selfing and wonder where the talk is. I did post a photo of a wee grandchild playing while tv playing and three other in room were on their gadgets. The discussion was good. No answers. But, the discusses all recommend Lahey's book.
In a period when helicopter parents strive for kids and overpower them, success comes without self motivation, and absent of failing
, or at least the skills needed to overcome failure, grit and resilience and desire. Good to,learn early that they are part of a community and tasks are performed not for bribes or pay, but part of all pulling together.
She spends a few pages on bullying and how to deal with it. Certainly for sandbox or recess in early years letting kids figure out how to work together through squabbles and fights is good. How do we handle the issues when kids get older? Teddy Roosevelt said learn to box. Sometimes really big kids pummel the weak. Stepping in may be needed.
Bullying which now is felt a larger part of school with snubbing, nasty notes, cliques causes no bodily harm, but is part of life. Dealing with it
Is a difficult task for teachers and parents.
I applaud the new generation of teachers and parents who work as teams. A quarter century ago the moms at our kids elementary school were called the mafia moms. No prisoners. I recall an episode when well dressed ladies said whatever the vogue n radon gas wrong, ..they knew cause they could read and they didn't have to learn what their kids were saddled with and the new math, now very old, was wrong. Maybe now teachers and parents can complain together about the evil core, or how much time the teachers should spend prepping for a test.
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Reviewed in the United States on January 16, 2016
★★★★★ 4
Good tips, though possibly over-obvious to some...
Format: Kindle
if you're prone to be a helicopter parent. If you're not, then I would say read it anyway so you can see where other parents might be approaching parenting. It pretty much fits in with other parenting books I've read such as "How Children Succeed" by Paul Tough, and "Grit" by Angela Duckworth.
As with any parenting book, how useful you find it depends on what you're already doing. One of the examples that the author uses in the book is that of a kid forgetting his/her lunch at home. If you "rescue" the kid by driving his lunch to school, then there's no motivation for him to remember it in the future. Got it! When I was a kid and I forgot my lunch at home, I was not then able to concentrate at school for fear of my mother's reaction when I got home. She needed to "teach me a lesson" by shaming me for forgetting it, cussing me out, turning it into a moral issue, and giving me the silent treatment for several days. When that's the consequence for forgetting a lunch, then going hungry for a day seems trivial. In fact, it would be less stressful to simply declare that I'm no longer bringing lunch to school anymore so that I can't possibly forget it again, and protect myself from a bipolar parent's mood swings. But some parents can't even let their kids go hungry for the day, even though suffering the consequence's of one's behavior doesn't even feel like punishment. If you had a parent like mine, you would hardly need to read this book, and yet I would recommend it so you can understand what other people's parents are doing.
My kid is 2 now. I pretty much let him learn from experience, although I outright ban dangerous and unhygienic behaviors. This book pretty much reinforces my approach anyway.
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Reviewed in the United States on August 2, 2017
★★★★★ 5
Infant favorite
Format: Board book
Love the colors, very sturdy.
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Reviewed in the United States on May 27, 2026
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