c*******4 发帖数: 41 | 1 My protein is around 20kDa, and I see many degraded protein bands (from
10kDa to 3kDa) on the gel. I tried to wash my protein several times using
the amicon centrifugal filter at 10kDa cutoff, but I still see those
degraded protein bands on the gels. I also add proteinase inhibitor and also
do the purification on ice to prevent the degradation, but it does not help
. Does anyone know what is the best way to purify or get ride of these
degraded protein bands on the gel? thanks. |
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b*******r 发帖数: 179 | 2 well, if you are doing crystallography then 20kda protein is fine
unless the part that's getting degraded is something they're interested in
looking at
if you wants to save it, i would try coexpressing the protein with a
binding partner
that binds the region that's getting degraded
hopefully the binding partner can stabilize it
so that it won't get degraded |
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a***n 发帖数: 578 | 3 thank for your information.
The problem I try to consider is target polymer will degrade at plantinum
surface under oxygen atomsphere. My initial idea is that the oxygen dissociate
on the plantinum surface as radical. Then this radical attack the side chain
of polymer and then degrade. Is it reasonable mechanism?
Are there any book to talk about degradation? Thanks a lot. |
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F***Q 发帖数: 6599 | 4 http://chargedevs.com/newswire/model-s-fan-builds-database-of-b
about 5% capacity loss after 60k miles (85k km), not so bad
Edit: the conclusions are: "The collected data indicate that the rate of
degradation starts out relatively fast, then slows down as more mileage is
put on the car. After about 50,000 miles, the overall range degradation is
about 6%. From that point onwards, range is lost at a rate of approximately
1% per 30,000 miles."
Edit2: original blog post URL
http://steinbuch.wordpres... 阅读全帖 |
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T**********t 发帖数: 1604 | 5 看RNA degradation好像一般都是用agilent的bioanalyzer比较准,用量很少,结果快
而且直观,就是他家的chip和仪器比较finicky,操作起来要很小心,否则一点小纰漏
就废掉一块chip。
如果是细胞里提的总RNA,bioanalyzer的分析软件能给出RIN数据,叫RNA integrity
number,数字越低,RNA降解得越厉害。
但我估计你看的RNA是short RNA oligo library,可能不会有RIN吧,我没试过。但是
你的sample如果用analyzer跑一下,出来的结果可以像胶一样看,有没有degradation
应该很容易看到吧。只需要几分钟的时间,有条件的话,试试看吧。 |
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s******y 发帖数: 28562 | 6 有啊。
Proc Natl Acad Sci U S A. 1995 Nov 21;92(24):11259-63.
Signal-induced degradation of I kappa B alpha requires site-specific
ubiquitination.
Scherer DC, Brockman JA, Chen Z, Maniatis T, Ballard DW.
Source
The inhibitor protein I kappa B alpha controls the nuclear import of the
transcription factor NF-kappa B. The inhibitory activity of I kappa B alpha
is regulated from the cytoplasmic compartment by signal-induced proteolysis.
Previous studies have shown that signal-dependent phosphorylation ... 阅读全帖 |
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s******s 发帖数: 13035 | 7 你说体内么?
是不是degrade和function没啥直接关系吧。misfold了就容易degrade,
这完全和function是间接关系。你的啥gain或者loss完全是遗传学的概念,
要看分子层面到底怎么回事
? |
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m**c 发帖数: 2103 | 8 60,000 miles = 96,560 km
原文5万mile写的很清楚啊。 After about 50,000 miles, the overall range
degradation is about 6% |
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g******z 发帖数: 5809 | 9 after 2 years, will degrade to regular USB Flash Dirve's speed |
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c********n 发帖数: 1577 | 10 我最近买了3个电源,什么evga,antec,thermaltake
可是还没有装机玩。。我要回国一阵子了,半年至少不会来
我记得以前有个哥们跟我说过,电源不能放,放个1年半,就开始电容什么的degrade
是真的么、 |
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c*******o 发帖数: 8869 | 11 if you do IP, you will see heavy chain around 50kd. If it is straight western
blot of tissue lysate, maybe the 50kd band is heavt chain of antibody from
tissue? also, RTK degradation is quite common thing. |
|
w******e 发帖数: 1187 | 12 RNA有OD260,NTP也有OD260,那RNA被degrade的过程OD260会有改变吗?
如果没有的话,有什么其他好办法能不用run gel就看出RNA的integrity呢?
多谢! |
|
a***e 发帖数: 1010 | 13 if OD260/280 is bigger than 2.2, there is a possibility of degradation. |
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s***m 发帖数: 6197 | 14 看是什么样的degradation了
一般260/280不是很能说明问题
如果是cleavage的话,还是跑胶比较好
如果是降解成mononucleotide
跑个TLC plate,带正点的那种
理论上也可以吧
可能就是量要多一点 |
|
|
h********n 发帖数: 4079 | 16 这个我以前用过, 很爽的.
degradation |
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m****M 发帖数: 360 | 17 Of course not. I got perfect ratios of my RNA samples from nanodrop, but when
I checked them on bioanalyser, they were all degraded (around 7 or lower).
A simple example, you can get perfect ratio when you measure DNA primers. I
have done a lot RNA work, trust me or not? It depends on yourself. -:) |
|
T**********t 发帖数: 1604 | 18 就是2100。我们实验室这台好久没人动了呢。:)
Bioanalyzer有很多应用啊。你们实验室开发microfluidics的,说不定可以借鉴一下,
或者在它的基础上开发点别的应用什么的,总之别跟他说你就是为了跑RNA看
degradation,不然他肯定让你自个儿跑个胶就完了。 |
|
C*******e 发帖数: 4348 | 19 agilent的bioanalyzer现在用的很多
的确又快用量又少
但是说白了其实也就是毛细管胶
一样是跑胶
一样是看rRNA的比例
没有这个条件的其实跑个琼脂糖电泳
照个照片
然后选择显示migrate/intensity
出来的图是一样一样的
只是没有人那样计算“RIN”而已
degradation |
|
s****z 发帖数: 50 | 20 Anyone has idea on how is miRNA degraded? What enzyme(s) are involved in
this process?
Thanks! |
|
b******9 发帖数: 102 | 21 read these papers:
1.Degradation of microRNAs by a Family of Exoribonucleases in Arabidopsis.
Science
2.Active turnover modulates mature microRNA activity in Caenorhabditis
elegans. Nature
3.The widespread regulation of microRNA biogenesis, function and decay.Nat
Rev Genet. |
|
a*****x 发帖数: 901 | 22 控制expression或者activity的很多,但是有控制degradation的吗? |
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r******k 发帖数: 446 | 23 大神们, gain of function的mutant protein会不会比WT被降解的更快呢?以我所知
,一般不都是loss of function的mutant form of protein会被degrade更快么??? |
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r******k 发帖数: 446 | 24 我一般的理解是mutant 更稳定。 比如p53的mutation比 p53 WT更稳定,cause更多的
bad effect。
但是对于我这个我还真没想明白,如果mutant protein能degrade更快,能说明啥呢? |
|
c******y 发帖数: 193 | 25 Thank you all for the reply!
1) Which amino acid in the peptide should be changed to d-AA? or any amino
acid change could prevent the peptide degradation.
2) Cyclization means make the whole peptide be cyclic, or just part of it?
Are there any reference on the cyclization protect peptide?
3) thanks for sharing peptoid! Are there any review on this?
Thank you again! Good night, sleep tight! |
|
T****O 发帖数: 407 | 26 1. Make it all d-AA with the reversed sequence, a.k.a., Retro-Inverso
2. Whoever saying cyclization can prevent degradation is ... anyway. It's
easy to try and not expensive.
3. Peptoids are very different. Don't branch out too much.
Finally, are you sure your peptides were broken down by enzymes in vivo?
How did you analyze or track peptides in vivo? |
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c*********o 发帖数: 2 | 27 请问各位做光伏太阳电池的高人,诸位做 acceleration degradation test 都用什么
equipment呀?
非常感谢!!! |
|
a***n 发帖数: 578 | 28 what does this termology mean?
what are the reasons causing degradation?
Thanks |
|
a***n 发帖数: 578 | 29 Can a metal catalyst degrade the polymer? by what mechanism.
Let us say plantinum.
What is the mechanism for 热降解?
Thanks. |
|
d****n 发帖数: 237 | 30 你去看看文献吧,这些我就不懂了,爱莫能助。
书要你自己去你所在学校的图书馆里去查,
我知道有本杂志,Polymer Degradation and Stability。
dissociate |
|
a***n 发帖数: 578 | 31 en. just found several good papers. some papers talked about oxygen dissociate
on Pt surface and produce OH. and O2H. radicals and attack alfa carbon to
degrade the polymers. partially as I assumed.
chain |
|
p******g 发帖数: 92 | 32 I'm looking for inexpensive bio-degradable polymers that can form viscous
solutions when dissolved in water. I want to suspend some inorganic particles
in water. can you give me some suggestions? I heard that some surfactants can
also be used to make viscous solutions. Is that true? what are some
surfactants that are commonly used? thank you in advance. |
|
w********h 发帖数: 12367 | 33 can we get both fair mechainical property and degradation rate? |
|
p******g 发帖数: 92 | 34 how to inhibit their degradation at high temperatures, say 150oC. dilution
solution in water. |
|
P******n 发帖数: 1220 | 35 Such as lifetime behavior of MEMS due to material degradation/aging and vary
multi-physical environment? Any useful papers on this topics? Thanks. |
|
c*********o 发帖数: 2 | 36 请问各位做光伏太阳电池的高人,诸位做 acceleration degradation test 都用什么
equipment呀?
非常感谢!!! |
|
|
|
t*****y 发帖数: 445 | 39 Summary of the Amazon EC2 and Amazon RDS Service Disruption in the US East
Region
Now that we have fully restored functionality to all affected services, we
would like to share more details with our customers about the events that
occurred with the Amazon Elastic Compute Cloud (揈C2 last week, our
efforts to restore the services, and what we are doing to prevent this sort
of issue from happening again. We are very aware that many of our customers
were significantly impacted by this event, and as ... 阅读全帖 |
|
r****y 发帖数: 263 | 40 收入申请书模板类别http://user.qzone.qq.com/2915799706/blog/1398824866
Petition letter (参照laoda555超长经验和hotmale的NOID回复写的):
XX , 2010
U.S. Department of Homeland Security
Citizenship and Immigration Services
RE:Applicaions: I-140 Immigration Visa Petition
Petitioner: Dr. B AA
Beneficiary: Dr. B AA (AXXXXX)
Category: Alien of Extraordinary Ability
Dear Immigration Officer:
The following is respectfully submitted in support of my petition for
classification as a qualified immigrant under the First Preference ... 阅读全帖 |
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x****u 发帖数: 530 | 41 谢谢大家的回复。
下面这篇文章是说GFP half life是2.8h吗?
Automated live cell imaging of green fluorescent protein degradation in
individual fibroblasts.
Halter M, Tona A, Bhadriraju K, Plant AL, Elliott JT.
Source
Abstract
To accurately interpret the data from fluorescent proteins as reporters of
gene activation within living cells, it is important to understand the
kinetics of the degradation of the reporter proteins. We examined the
degradation kinetics over a large number (>1,000) of single, living cells
from a... 阅读全帖 |
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f**d 发帖数: 768 | 42 这是一本计算神经科学的优秀著作,全文拷贝这里(图和公式缺),有兴趣的同学可以
阅读
如需要,我可以分享PDF文件(--仅供个人学习,无商业用途)
From Computer to Brain
William W. Lytton
From Computer to Brain
Foundations of Computational Neuroscience
Springer
William W. Lytton, M.D.
Associate Professor, State University of New York, Downstato, Brooklyn, NY
Visiting Associate Professor, University of Wisconsin, Madison
Visiting Associate Professor, Polytechnic University, Brooklyn, NY
Staff Neurologist., Kings County Hospital, Brooklyn, NY
In From Computer to Brain: ... 阅读全帖 |
|
Z*****o 发帖数: 4090 | 43 http://www.stereophile.com/features/368/index.html
By Robert Harley • Posted: Nov 1, 1993
Not that long ago, digital audio was considered perfect if all the bits
could be stored and retrieved without data errors. If the data coming off
the disc were the same as what went on the disc, how could there be a sound-
quality difference with the same digital/analog converter? This "bits is
bits" mentality scoffs at sonic differences between CD transports, digital
interfaces, and CD tweaks. Becaus... 阅读全帖 |
|
w***7 发帖数: 1637 | 44 宋的文章够了,experience不够让人信服。
宋的博士学位在中科院拿的,在us做了3-4 年postdoc。宋在us 待的时间太短,对美国
博士培养体制和科研体制先进之处了解得可能不够。一同面试院长的有几个candidate
文章比宋好。武大可能是看到宋在中科院当个官,搞个管理就把it请过来了。武大应该
找一个在美国拿个PhD,然后在us 做postdoc后海归的当院长。
宋自己的科研做得很好,但对宋领导一个院做出像样的改革,大幅提高整个院科研水平
不太看好。
http://www.bio.whu.edu.cn/News_reads.asp?cid=174&id=2574
学习经历:
1993-1997 南京大学生物科学与技术系,学士
1997-2002 中国科学院上海生命科学研究院生物化学与细胞生物学研究所,博士
主要工作经历与任职情况:
2002-2005 美国德克萨斯大学西南医学中心,博士后
2005-2014 中科院上海生命科学研究院生物化学与细胞生物学研究所,研究组长,研
究员,博士生导师,“百人计划”择优支持,“百人计划”终期评估优秀。分子生物学
国家重点实验室... 阅读全帖 |
|
w***7 发帖数: 1637 | 45 宋的文章够了,experience不够让人信服。
宋的博士学位在中科院拿的,在us做了3-4 年postdoc。宋在us 待的时间太短,对美国
博士培养体制和科研体制先进之处了解得可能不够。一同面试院长的有几个candidate
文章比宋好。武大可能是看到宋在中科院当个官,搞个管理就把it请过来了。武大应该
找一个在美国拿个PhD,然后在us 做postdoc后海归的当院长。
宋自己的科研做得很好,但对宋领导一个院做出像样的改革,大幅提高整个院科研水平
不太看好。
http://www.bio.whu.edu.cn/News_reads.asp?cid=174&id=2574
学习经历:
1993-1997 南京大学生物科学与技术系,学士
1997-2002 中国科学院上海生命科学研究院生物化学与细胞生物学研究所,博士
主要工作经历与任职情况:
2002-2005 美国德克萨斯大学西南医学中心,博士后
2005-2014 中科院上海生命科学研究院生物化学与细胞生物学研究所,研究组长,研
究员,博士生导师,“百人计划”择优支持,“百人计划”终期评估优秀。分子生物学
国家重点实验室... 阅读全帖 |
|
a***k 发帖数: 1038 | 46 【 以下文字转载自 paladin 讨论区 】
发信人: atack (小军号), 信区: paladin
标 题: 红死病 by 杰克.伦敦
发信站: BBS 未名空间站 (Sat Oct 11 07:40:15 2014, 美东)
I
The way led along upon what had once been the embankment of a railroad. But
no train had run upon it for many years. The forest on either side swelled
up the slopes of the embankment and crested across it in a green wave of
trees and bushes. The trail was as narrow as a man's body, and was no more
than a wild-animal runway.
Occasionally, a piece of rusty iron, showing through ... 阅读全帖 |
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F***Q 发帖数: 6599 | 47 #1:
incorrect, PHEVs can travel long distance without recharging
#2:
incorrect too, nearly all EVs (BEV/PHEV) can be charged from a household
outlet (that's called Level 1/L1 charging) that is abundant in any garage.
#3:
it is not unknown anymore. for several models, it has been long enough that
the lifespans of different batteries can be tested by both the manufacturers
and the existing cars on the road. yes there is degradation, but it is less
than what people had speculated
so far temperature... 阅读全帖 |
|
a***o 发帖数: 54 | 48 http://kidjacked.com/legal/spanking_law.asp
Alabama
[The Code of Alabama 1975]
Parent/guardian/person responsible for care and supervision of a minor/
teacher or other person responsible for care and supervision of a minor for
a special purpose may use reasonable and appropriate physical force when and
to the extent he reasonably believes it necessary and appropriate to
maintain discipline or promote welfare of the child.
§ 13A-3-24. [Criminal Code]
[See also: Endangering welfare of child. Secti... 阅读全帖 |
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m*****i 发帖数: 4342 | 49 NEW YORK (TheStreet) -- Imagine a company that has predicated its future on
building out large-scale projects in desert conditions. Now imagine the
technology the company uses doesn't have a long-term track record of
performing under intense heat.
You don't have to imagine the company. It's First Solar(FSLR), and based on
the revelation in its Tuesday conference call with analysts that high-
temperature degradation issues have surfaced with its panels, it's the most
alarming prospect for a compa... 阅读全帖 |
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c*********o 发帖数: 1734 | 50 我可以把你推荐给editor,感兴趣的话,尽快PM我。
你需要把你的简单背景发给我, 包括名字,email,现在的organization和title。 还
要在这个journal的reviewer database注册一下。
Title: Thermally induced degradation pathways of three different antibody-
based drug development candidates
European Journal of Pharmaceutical Sciences
ABSTRACT:
Protein-based medicinal products are prone to undergo a variety of chemical
and physical degradation pathways. One of the most important exogenous
stress condition to consider during manufacturing, transport and storage
pro... 阅读全帖 |
|