w***r
发帖数: 709 | 1 这个人太牛X了。他在song h 甲基化文章真的很炫!！
但是我个人觉得
Cytochrome P450 drives a HIF-regulated behavioral response to reoxygenation
by C. elegans. Science (2013) Vol. 341 no. 6145 pp. 554-558. PMID: 23811225
这篇文章似乎新意不够，hif 和p450和缺氧的关系早就知道了。下面是一篇，还有不少
有明白人讲一讲吗？
Inhibition of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)-stimulated Cyp1a1
promoter activity by hypoxic agents.
Kim JE1, Sheen YY.
Author information
Abstract
Since hypoxia-inducible factor-1alpha (HIF-1alpha) and the arylhydrocarbon
receptor (AhR) shared th... 阅读全帖 |
|
w***r
发帖数: 709 | 2 这个人太牛X了。他在song h 甲基化文章真的很炫!！
但是我个人觉得
Cytochrome P450 drives a HIF-regulated behavioral response to reoxygenation
by C. elegans. Science (2013) Vol. 341 no. 6145 pp. 554-558. PMID: 23811225
这篇文章似乎新意不够，hif 和p450和缺氧的关系早就知道了。下面是一篇，还有不少
有明白人讲一讲吗？
Inhibition of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)-stimulated Cyp1a1
promoter activity by hypoxic agents.
Kim JE1, Sheen YY.
Author information
Abstract
Since hypoxia-inducible factor-1alpha (HIF-1alpha) and the arylhydrocarbon
receptor (AhR) shared th... 阅读全帖 |
|
g**********y
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aafes-thanks... 阅读全帖 |
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A******y
发帖数: 2041 | 4 Btw, I made a mistake, the name should be EC50 (effective concentration).
LD (lethal dose) is usually used in vivo for animal studies.
This is how my lab does EC50: We do 12 point titration either (1/2 or 1/3)
dilution per dilution. For example, for single digit microM compounds, we
do 200 micorM, 100 micorM, 50 microM, for 12 (Log[concentration] will be
your X-axis) points...After that we determine the viability of the cells (Y
axis). Then, you use a non-linear fit program (Prism is the mos... 阅读全帖 |
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A******y
发帖数: 2041 | 5 Depending on the target, you can have single digit microM (tens microM is
too high for a lot of targets) inhibitor if the target is novel. In my
field, you need around 100 nM and selective to the isotype to be able to
publish. |
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A******y
发帖数: 2041 | 6 There is no way 5 microM drug can be specific. Specificity can only be
gained through potency. It could be selective for a specific set of
isozymes but definitely not specific at 5 microM. Single digit nanoM to
picoM inhibitors could be very specific if you used at the correct
concentration.
target |
|
A******y
发帖数: 2041 | 7 For number 1, if you are worry about reviewers, you have to do two things: 1
. Mass spec will tell you where the modification is. 2. For your gene of
interest, you can use different but well know histone acetylation or
methylation marker doing chromatin IP. Both require experience and special
training to do them right though. Even if you can find specific actylated
ab for the resiude, reviewers will ask for these two experiments for a good
journal. As for as I know, histone H3 K9 Chip is ve... 阅读全帖 |
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A******y
发帖数: 2041 | 8 If it has certain absorbance or florescence that you know, you can use that
to determine the solution concentration. A drug that can only dissolve in
microM concentration in DMSO, it is a useless drug. You won't be able to
get into human at all though. |
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A******y
发帖数: 2041 | 9 The correct name is LD50 or ED50, but the mathematical equation is the same.
I hate to tell you that you don't have enough point to determine your LD50
. You need at least two log scale, but it look like your LD50 is around 50
(unit?). If it is microM, you are wasting your time. |
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A******y
发帖数: 2041 | 10 Most people look at GSH depletion because the cells already have a lot of it
. Some cells if you treated them with high glutamate, it will block
cysteine uptakes and deplete its GSH level. Your probe has a Micheal's
acceptor, so I you will have toxicity around 10s microM, but it is probably
okay to do short term live cell imagining.
I wounder if you treated with certain compounds, would you be able to
determine specific GSH pool depletion because there is a cyto- and a mito-
pool of GSH.
Tsie... 阅读全帖 |
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b**********n
发帖数: 34 | 11 Using the solution stablized with 1 microM melamine to assay the melamine
with the concentration in nanoM range. This
really likes using buffer solution to measure acid and base in solution.
This is strong evidence to claim that authors fictionalized their data. |
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k****o
发帖数: 728 | 12 我实在弱酸pH4条件下做的，peptide浓度只有0.5 microM. 而且，MS没有显示有大分子
量的产物。 |
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A******y
发帖数: 2041 | 13 Tell me about it...I have nightmare about publishing...found a novel
selective inhibitor and only inhibits one out of 11 isoenzymes (I did IC50s
on all of 11 isozyes on all compounds), the reviewers still think it is not
specific because its IC50 is at ~1 microM. I even over-expressed the
proposed target and shown the over-expression attenuates the toxicity of the
compound...still under review :( over a month. |
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J*J
发帖数: 132 | 14 主要是1 microM， 如果是1 nM， 可能你不会遇到这个问题了
IC50s
not
the |
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