p*****n
发帖数: 981 | 1 有好的ligand么，作一下titration? |
|
w******y
发帖数: 2504 | 2 Could anybody send me a copy of this paper:
Kinsey CG, Bussolati G, Bosco M, Kimura T, Pizzorno MC, Chernin MI, Cassoni
P, Novak JF. Constitutive and ligand-induced nuclear localization of
oxytocin receptor. J Cell Mol Med. 2007 Jan-Feb;11(1):96-110.
My email address is: p***********[email protected]. Thanks so much. |
|
D******9
发帖数: 2665 | 3 I would like to study the cell signaling pathway initiated by a cell surface
ligand. However, I do not know its receptor. What is the best way to find
its receptor? Any good ideas? Thanks a lot. |
|
p*****m
发帖数: 7030 | 4 1 你google一下从EPO receptor到serotonin receptor到TRP channel怎么搞出来的
就行了 简单来说就是搞一个没有receptor的cell line，把有receptor表达的cDNA
library转进去 然后加ligand，看下游的phenotype就行了 可以是binding activ
ity，也可以是downstream signaling,等等
2 没有 但是如果你有个范围可以大概用agonist/antigonist猜猜看
specific |
|
p*****m
发帖数: 7030 | 5 你是什么性质的ligand? 也不是不行 但是实际上会很困难 |
|
s******y
发帖数: 28562 | 6 There is back-ward and forward screening methods
For forward screening, you can do crosslinking and pulldown of candidate
proteins, and then send the products for mass spec
For back-ward screenings, you can use random KO library (usually with yeast)
to select the clones that are resistant to your ligand (with proper
screening assay), and then figure out what gene is required for the signal
pathway.
Either way it is not a small work
surface |
|
w******e
发帖数: 1187 | 7 competition binding, 2 ligands, 2 receptors. any recommendation on
equations? |
|
D*a
发帖数: 6830 | 8 你要看看你exon3是干什么用的，多少bp
比如如果是膜蛋白，是不是胞外亲和ligand的位点，或者往胞外运输的信号，或者跨膜
部分，或者下面的kinase部分正好能被你切掉。
而且最好是少了exon3之后能造成exon4的移码突变，然后弄出几个stop来。
这样才能确保你真能ko掉
完了你得在老鼠上测测全长的mrna是不是真的下降了。 |
|
H*****e
发帖数: 120 | 9 Everytime, it is amazing to see the 3-D binding model for small molecule
binding a protein. Now I found a small molecule (a boring one!) binding a
protein (with known crystal structure). For publication purpose, I am very
interested in try to have such a model because I noticed such a computation
based modeling is publishable. Although I am not a structure person, I was
told that it can be done by computation based prediction in a seminar. I
tried google but found too many and too confusing. |
|
m*****o
发帖数: 17 | 10 If you put me as one of the co-authors, I will do it for you. |
|
H*****e
发帖数: 120 | 11 I am open mind for it. Drop me your contact information in my box and we
can discuss it. Your contribution (I believe this is the right term for
authorship) shall be recognized. In addition, I care more about how to do
it and very interested in learning it (even just for fun although I have no
idea when it will be next time that I want to do this kind of work). Teach
me how to do it is also your contribution. |
|
m*****o
发帖数: 17 | 12 It is fine. I agree with you about the "contribution" term.
Please check your mailbox. |
|
w******e
发帖数: 1187 | 13 单抗通常是用来deliver别的东东的targeting ligand呵呵。
说起来gene therapy/antisense惨淡收场之后，siRNA/microRNA能又这么快的
火起来，可见钱多的用不完，总得搞出个概念炒一炒呵呵。
真正的成果，我现在缺乏expertise去判断。什么是检验标准呢？
我觉得得是commercialization。可是aptamer 6年前就有药了，
nanoparticle 15年前就有药了。按说也prove了concept吧？
可是接下来又是长期的沉寂。 |
|
w******e
发帖数: 1187 | 14 Thank you very much for the information. I'm mainly interested in
nanoparticles for in vivo application. a desired one would be a.
biocompatible,
b. controlably releasing, c. easily modified w/ targeting ligand & drugs,
d. do not require expensive instrumentation or lengthy prep, e. has long
shelf life, f. consistent in size & shape, g. has minimal loss in RES.
but really anything that satisifies c & d would work:) Could you pls
recommend some papers/protocols for me? as long as I can easily sti |
|
q*****n
发帖数: 331 | 15 同意楼上的意见。如果你只有生物的背景，没有化学合成的背景，最好的办法是找个做
Nano的lab合作。
满足下列条件的：
c. easily modified w/ targeting ligand & drugs,
只有Invitrogen的qdots (commecially available). 但是，他家的qdots
accumulate non-specifically而且不能穿越血管到达肿瘤。
前两年有不少用Invitrogen的qdots做in vivo imaging 的，但是现在再用的话恐怕很
难发文章的。因为他的缺点大家都知道了。
找个做nano的lab, 即使他们的nano particle 也有这些毛病，但是不广为人知的话，
还能发文章的。
个人感觉现在进nano已经不是好时候了。
欢迎有不同意见的一起讨论。 |
|
T*****u
发帖数: 3257 | 16 Postdoctoral fellow position in Laboratory of Cardiovascular Science,
National Institute on Aging, NIH.
Research program:
Receptor for advanced glycation end products (RAGE) is a pattern recognizing
receptor that interacts with multiple ligands and elicits cellular
inflammatory responses. Signaling via RAGE has been implicated in several
chronic, aging related illnesses including atherosclerosis, diabetes, and
Alzheimer’s disease. Persistent inflammation has been identified as an
important physi |
|
w******e
发帖数: 1187 | 17 我看一个methods上，在incubation结束后狂dilute一下，模拟ligand无法rebind的
状态，那么complex的数量根据时间成指数衰减，把几个time point上的
value plot一下可以搞出个koff来。
ITC倒没听过，不过0.1~0.2mg...我的蛋白都是买的，50ug300大刀，肯定没得玩~ |
|
D*a
发帖数: 6830 | 18 还有要看你们当时是怎么决定的KO的strategy吧
像我们的老鼠敲掉的是跨膜蛋白的ligand结合位点的exon，然后敲掉之后在下一个exon
中造成移码突变，然后造成几个stop，这样就确保蛋白质没有作用。
还有我们的引物设计是这样的
引物a －－loxp－exon－－引物b－－loxp－－反向引物c
这样wt和lox是靠bc来区别，ac相比之下太长扩增效率很低，ko之后就是ac扩增，这样
wt lox excised都可以用这三个引物来区分。我觉得这种pcr比你的好些，要不你自己
重新设计引物试试吧。 |
|
|
|
t*********1
发帖数: 418 | 21 我不确定这个受体是不是autoreceptor， 目前还没有太多有关这个受体的信息。但是
我知道有几个神经元上表达这个受体。如果是autoreceptor的话，会怎么样？不太懂
autoreceptor, 能给具体解释一下吗？谢谢！ |
|
t*********1
发帖数: 418 | 22 查了一下AUTORECEPTOR的定义，我认为我研究的受体不应该是AUTORECEPTOR。 |
|
|
y****i
发帖数: 2194 | 24 一般来说 antagonist的作用是neutral的
如果有相反的作用的话 一般叫做inverse agonist
dr
li
binding
叫做
b
高
这时 |
|
b*****l
发帖数: 9499 | 25 多谢！翻出教材来又查了查，的确如此。
analog/
种
引起
affinity |
|
n***w
发帖数: 2405 | 26 是叫partial agonist。
dr
li
binding
叫做
b
高
这时 |
|
S*****s
发帖数: 287 | 27 同意。理论上来讲 antagonist 的作用是结合到 receptor 的 binding pocket 上让
agonist 不能结合，本身并不能抑制 receptor 的活性。 |
|
w***a
发帖数: 4361 | 28 NR算transcription factor的一种吧，一般需要ligand激活才入核bind DNA。 |
|
s***o
发帖数: 1189 | 29 Thanks for all the informative posts above.
I've been working on receptor-ligand interactions for a while and will dive
in job market pretty soon. Lately I've found quite a few openings in China
for HTS ppl, from CROs and big name pharmas as well.
Yet the question is, if continuing our career in the bio-tech field, would
those Chinese-based companies provide a decent environment for us to grow?
I've heard ppl saying the core of R & D stays here and work assigned to
Chinese company is basically l... 阅读全帖 |
|
h***a
发帖数: 145 | 30 that knockout of A increases the activation of B by specific ligand C. But
adding back A into the knockout does not reduce the activation of B to the
normal level (actually no influence).
百思不得其解啊。 |
|
s****9
发帖数: 932 | 31 Another alternative to consider is Flt3-ligand blockade. It remarkably
reduces DCs numbers, but you need to check more reference to see whether it
also influence some other myeloid compartments. |
|
z*******6
发帖数: 679 | 32 嗯，两年前学免疫的时候老师就是免疫组学。。。天天说，当时一直没搞懂到底啥是组
学。。。
嗯，现在看上的两个免疫实验室一个是做HLA的，这个感觉就是在测序，找序列，找
ligand，感觉学不到很多免疫相关的技术，优点是有钱，课题比较intriguing，貌似是
个潜力课题，缺点是技术上学不到太多东西，他的系统都很成熟了，还有就是老板明确
说了他把握实验进展方向，不能有太多偏离的想法，告诉我说PhD就是要学会如何focus
；实验室氛围很好，很热闹，老板也很好，尤其对学生好。。。
另一个是做宿主对HSV的免疫防御的。。。整个技术都是免疫常用的，in vitro in
vivo的都能做，但感觉实验室貌似今年没钱了，但他自己号称还有？（这个是听现在在
他实验室rotation的同学说的），然后就是是小实验室，还有就是觉得课题没太大潜力
，这个还没有去rotation，下一轮可能去看看，具体相处不相处的来还是另外一回事了
。。。 |
|
T**********t
发帖数: 1604 | 33 有，叫splinted ligation。
因为T4 DNA ligase的功能是特异性修复双链DNA上的nick。所以需要设计一个splint
oligo，和你需要ligate的两个oligo的两端各自互补。像这样：
图里显示的是连接RNA，不过DNA也是一样的。
这个protocol在这里：
http://www.megaupload.com/?d=QT50D23R
[7] Joining of RNAs by splinted ligation
Melissa J. Moore and Charles C. Query
Methods in Enzymology
Volume 317, 2000, Pages 109-123
RNA - Ligand Interactions, Part A |
|
m**z
发帖数: 787 | 34 1. in-gel tryptic digestion followed by MS or MS-MS
2. ESI-MS or functional assay
3. CD denaturation
4. cys oxidation and others? not quite sure, guess depends on the mass
difference
5. not sure... MS-MS?
6. increase salt, change buffer condition, lower temperature, add a ligand
to stabilize, etc. |
|
N*****h
发帖数: 324 | 35 EGFR表达是epithelial细胞的特征之一，为什么用EGFR的ligand (EGF, TGF-beta)反而
能诱导细胞往mesenchymal那边呢？要做rotation选课题，这个问题实在没想明白。谢
谢各位大牛。 |
|
p******i
发帖数: 1092 | 36 pubmed裡面有……
而且別人已經用這個當ligand 發表過文章了…… |
|
a****m
发帖数: 693 | 37 Mechanisms of Development
Volume 127, Issues 7-8, July-August 2010, Pages 358-370
The interaction between two TGF-β type I receptors plays important roles
in ligand binding, SMAD activation, and gradient formation |
|
|
q*****n
发帖数: 331 | 39 Insulin: insulin receptor
IGF-I: type I IGF receptor
The presence of the receptors in the tissue suggest that insulin or IGF-I
may stimulate their growth.
The binding of the ligands (insulin or IGF-I) will cause receptor
phosphorylation and activation, which in turn, activate downstream signaling
pathways, such as MAPK and Akt.
Use physiological concentration should be fine.
IGF-I: 5nM
insulin: 10-20 nM
If you want detailed information, search pubmed. There are so many reviews
outthere. |
|
s***o
发帖数: 1189 | 40 It is written in textbook.
but here is some hint, the identified receptors are mostly named according
to their noticeable ligands. |
|
F*******2
发帖数: 103 | 41 欢迎后面同学拍砖：最简单的方法就是把你的ligand covalent bind到什么resin上，
然后pull down一下吧。 |
|
s******y
发帖数: 28562 | 42 因为很多受体有clustering 的倾向。而且clustering 对它们的实际活性有很大的
关系。如果是到处散布的话，不能定点的激活某一个地方，会导致局部的signal
molecule浓度不够高，就会导致信号传播不下去。
具体的clustering 的机理对于不同受体可能不同，有的是依赖于对膜上某些脂分子
的结合特性，脂分子出现clustering 在先； 有的是依赖于受体分子活化变构后暴露出
来的互相结合的集团而实现clustering; 还有的是依赖于binding ligand 的
clustering. |
|
|
r****t
发帖数: 10904 | 44 Contributions
A.S. designed experiments and performed all the phenotypic characterization
of the mouse mutants, the cortical neuron culture, organotypic slice culture
rescue experiments and electroporation of cerebellar acute slice cultures.
A.A.-P. performed the initial biochemistry on the binding of ephrin B
ligands to Reelin, designed experiments, interpreted results and wrote the
manuscript.
这个 A.A.-P. 干系逃不掉啊。 |
|
r****t
发帖数: 10904 | 45 Contributions
A.S. designed experiments and performed all the phenotypic characterization
of the mouse mutants, the cortical neuron culture, organotypic slice culture
rescue experiments and electroporation of cerebellar acute slice cultures.
A.A.-P. performed the initial biochemistry on the binding of ephrin B
ligands to Reelin, designed experiments, interpreted results and wrote the
manuscript.
这个 A.A.-P. 干系逃不掉啊。 |
|
s***m
发帖数: 6197 | 46 文章1
链接：
http://www.nature.com/nchembio/journal/vaop/ncurrent/full/nchem
Molecular insights into the ligand-controlled organization of the SAM-I
riboswitch.
Heppell B, Blouin S, Dussault AM, Mulhbacher J, Ennifar E, Penedo JC,
Lafontaine DA.
Nat Chem Biol. 2011 May 1.
文章2
链接：
http://www.nature.com/nchembio/journal/vaop/ncurrent/full/nchem
Conformational capture of the SAM-II riboswitch.
Haller A, Rieder U, Aigner M, Blanchard SC, Micura R.
Nat Chem Biol. 2011 May 1.
麻烦发到l****[email protected]
或者提供下载链接
... 阅读全帖 |
|
m*****s
发帖数: 156 | 47 done
文章1
链接：
http://www.nature.com/nchembio/journal/vaop/ncurrent/full/nchem
Molecular insights into the ligand-controlled organization of the SAM-I
riboswitch.
Heppell B, Blouin S, Dussault AM, Mulhbacher J, Ennifar E, Penedo JC,
Lafontaine DA.
Nat Chem Biol. 2011 May 1.
文章2
链接：
http://www.nature.com/nchembio/journal/vaop/ncurrent/full/nchem
Conformational capture of the SAM-II riboswitch.
Haller A, Rieder U, Aigner M, Blanchard SC, Micura R.
Nat Chem Biol. 2011 May 1.
麻烦发到l****[email protected]
或者提... 阅读全帖 |
|
m******5
发帖数: 1383 | 48 最近作microarray数据（in the context of development ）
遇到几个比较有意思的CXCL family ，包括cxcl1, cxcl5等上调，下调
在网上查了查文章，似乎cxcl family ligand很多人都是在免疫学的context下研究的
，比较少看见在development biology的context下做的
所以想问问cxcl family在development里有什么研究成果？最好推荐一些好的综述，非
常感谢！！！ |
|
c********r
发帖数: 1125 | 49 这个chemokine CXCL family factors在发育里面的作用不是什么很新的东西了，在PGC
迁移，肌肉前体细胞迁移，血管发生上面都有报道了。
我记得比较清楚的是在斑马鱼里面的工作，你可以参考:
Siekmann, A.F., Standley, C., Fogarty, K.E., Wolfe, S.A., Lawson, N.D. (2009
): Chemokine signaling guides regional patterning of the first embryonic
artery. Genes & Development, 1;23(19): 2272-7.
Vasyutina, E., Stebler, J., Brand-Saberi, B., Schulz, S., Raz, E. and
Birchmeier, C. (2005)
CXCR4 and Gab1 cooperate to control the development of migrating muscle
progenitor cells.
Genes and Devel... 阅读全帖 |
|
b*******n
发帖数: 8420 | 50 LS说得没错，chemokine可以引导细胞迁移。
当然如果你非要联系到免疫方面的话（如果那几个发生变化的chemokine ligand/
receptor的主要作用是招募免疫细胞），那么一种可能的解释就是有些免疫细胞被招募
到某些site，消灭掉一些组织细胞，从而实现remodelling. |
|
