z*******w
发帖数: 79 | 1 +-------------------------------------------------------+
|BIND 8.2 - 8.2.2 *Remote root Exploit How-To* by E-Mind|
+-------------------------------------------------------+
(A) What is a DNS?
1. How do I query a DNS?
2. How do I find a vulnerable DNS?
(B) How do I edit DNS entries?
1. How do I find a Zone file?
2. How do I edit a Zone file?
(C) How do I exploit a vulnerable machine
1. What do I need to obtain before I could use the exploit?
2. Wha |
|
w********r
发帖数: 6 | 2 look at the code.
I compare it with the example program words by words.
and I have tried -lsocket -lsnl in linking.
(of course, the system says there is no such file or directory.
BTW, all the socket and bind are system calles. need to link
socket library?)
does not work at all.
I typed the error message, it says: the address is already
in use.
following is the code. thanks a lot!! |
|
m*****e
发帖数: 4193 | 3 So the answer is very clear: address is already in use. Try to bind a free
port. |
|
m*****e
发帖数: 4193 | 4 try telnet to that port you are trying to bind, and see whether you get
a Connection refused error. |
|
w********r
发帖数: 6 | 5 上次的问题已经找到原因了.
就是我试图要把几个socket都binding到同一个
地址(IP& port).
写的时候没注意... 就出了那种情况.
我想是应该有办法做到这一点的. seems apache did so.
有人有这方面的经验么?
恳请指点一二. |
|
m*******m
发帖数: 182 | 6 And SO_REUSEPORT.
You dont normally need bind in an UDP case. (It is
specified in the sockaddr when you call recv() or
recvfrom()) Multicast is a special case for UDP.
到: 】 |
|
p********n
发帖数: 82 | 7 啥叫binding theory啊？
需要的時候用反身，不需要的時候就不用唄 |
|
d*****u
发帖数: 17243 | 8 Sounds interesting
what do you mean by non-syntactic binding here?
paper? |
|
a***e
发帖数: 1010 | 9 what is HA control? something unrelated or related?
HA
input
binding |
|
|
a***e
发帖数: 1010 | 11 in these cases, both HA and IgG are background controls. Theoretically,
both of them should have no pulldown signals at all (= 0). If you divide
something by HA or IgG signal, any little variation will change your ratio
dramatically. So, neither of them is the right normalization.
A good normalization method is using 1% input as a control.
However, the best normalization is using some internal controls that are not
changed by your treatment. For example, your TF binds gene GAPDH and gene
X. |
|
a****o
发帖数: 1786 | 12 如果超声不充分的话，可溶蛋白含量少，不同细胞需要的超声条件可能不同
HA
input
binding |
|
e********r
发帖数: 147 | 13 在体外条件下，研究核糖体小亚基与mRNA的结合可以用ribosome toeprinting assays
这一类的方法。但在我们的研究体系里，由于调控因素可能较多，体外实验体系分析不
一定能够重现细胞内的条件，拿到positive实验结果的可能性较小。我想请问如果在细
胞内，有什么生物化学（或者遗传）的方法可以来检测核糖体与具体mRNA的binding活
性？ |
|
y******8
发帖数: 1764 | 14 how about Protein G B1 domain and binding motif on Fc |
|
w******e
发帖数: 1187 | 15 avidin-containing target + biotinylated probe + avidin-containing Ab,
看到了signal。可是只记得一个avidin能bind 几个biotin。莫非avidin
multimer会dissociate，and then interact w/ exogenous avidin? |
|
T**********t
发帖数: 1604 | 16 Avidin是鸡蛋清里提取的，是碱性的。
Streptavidin是链霉菌里提取的，是弱酸性的。
Neutravidin是化学修饰过的Avidin，是接近中性的。
Streptavidin和Neutravidin的优点就是non-specific binding比Avidin少。
不过你既然用的就是Streptavidin，那我就不清楚了。但是你的probe还是有可能跟
Streptavidin有cross-reactivity的。你screen probe之前用Streptavidin做过
negative selection了么？ |
|
w******e
发帖数: 1187 | 17 土问：啥叫4-15% gel啊？gradient？
实话说我对native gel跑蛋白一直没搞明白原理。是不是只有跟aptamer bind了
之后才会带负电才会migrate？那感觉应该比SDS的效果差非常非常多？刚才有
铜锈指出还能调gel pH，完全不知，窘了。。。 |
|
x**y
发帖数: 46 | 18 跟我最近的EMSA相似，给你一些有用的意见：
1. 如果你的蛋白有aggregation，加一些Triton100(0.1，0.5or1%). 听很多人说，这
很有用。
2. 用4-12% TBE gel （Invitrogen）。如果自己配，用4%。或者3%，下层加三分之一
15%防止free probe run out。
3. 如果还不行，用垂直配胶系统（Bio-rad配PAGE gel用的， 1.5mM）配Agrose胶（1%
？ 2%？记不清了）。里面可以加Triton & glycerol。
我的蛋白80kD， oligomerization 后500 or 1000kD （FPLC测定）。6%TBE gel完全在
well上，4-12%能进一点点，3%可完全进入但很难看。最近我们lab新来的Postdoc用了
Agrose胶，效果不错。我让他加了一些Triton & glycerol，效果更好了，binding
affinity 都增加了几倍。但具体浓度，我得问问他。 |
|
w******e
发帖数: 1187 | 19 又来请教了：话说我用filter binding assay测我aptamer pool跟target
protein的kd，测出的signal最高只有upper limit（probe不加protein不过
filter直接加到scintillation vial里的读数）的20％。。。虽说protein
理论上可能有一部分被wash off吧，但80％ loss也太离谱了吧。。。但
signal确实是plateau了，能fit出kd来（还挺好hiahia）。
请问牛人这种现象可能正常吗？btw我根本没敢多洗，加了mixture之后就
洗了1次1ml，不过neg control挺低的了。 |
|
w******e
发帖数: 1187 | 20 thanks a lot! Now I'm relieved:)
bind |
|
g*****n
发帖数: 241 | 21 我想做一个insulin-receptor binding assay，请问有没有什么protocol啊？我找了好
久都没找到
非常感谢！ |
|
d********r
发帖数: 211 | 22 体外养细胞的时候, 里面已经有些细胞自己分泌的ECM,
这时候如果加growth factor, 倒底是被细胞take up
还是bind to ECM? 尤其是某些有很强的特异吸附ECM的
growth factor. |
|
m*****n
发帖数: 760 | 23 有没有哪些网站，或者数据库，
能够比较靠谱的computationally predict转录因子的binding site？ |
|
|
I*****y
发帖数: 6402 | 25 make yourself clear first
binding |
|
m******5
发帖数: 1383 | 26 现在在对比同一个基因的人和老鼠的上游Promoter区域，想找潜在TF binding位点，请
问哪个网站最好用？ |
|
m******h
发帖数: 28 | 27 Did anybody know if the trans-acting factor binding consensus has direction.
For example, if the Smad3 recognizes the 5'CAGA3' sequence, can it
recognize the 5'TCTG3' sequence? Are there any experimental evidences? Thanks
for answering. |
|
C*******e
发帖数: 4348 | 28 想问问看版上做DNA binding protein的同学
有没有简单的方法，不做放射性同位素标记的，Mobility Shift Assay啊？
现在我们手头有一个细菌蛋白
是属于某个细菌细胞器的
该细胞器和DNA无关
但这个蛋白的具体功能未知
只是有不寻常的high pI (>11)
在纯化该蛋白的时候发现UV reading近乎DNA
事实也证明跑胶以后目标蛋白很少
UV reading一多半来自于DNA
过SEC之后能把蛋白和DNA分开
有没有办法做个简单的测试
验证该蛋白能结合DNA？
哪怕是非特异性的 |
|
t******6
发帖数: 1657 | 29 random binding selection |
|
s*********y
发帖数: 387 | 30 purchased a mAb against protein with domains of
A-B-C-D-E
The mAb is produced by recombinant protein of B plus a small region on A.
Now, I purified the GST-A, GST-B, GST-C, GST-D, GST-E, and GST-Full
length.
When testing binding by ELISA, only GST-Full length and GST-B can be
recognized by this antibody.
But, in SDS-PAGE and wester blot, GST-A, GST-B, GST-C, GST-D and GST-Full
length, but not GST and GST-E, can be recognized by this mAb.
Totally lost for an applicable explanation, please help. t... 阅读全帖 |
|
C*******e
发帖数: 4348 | 31 虽然我们也用pH 8的条件做GST蛋白纯化
但是好像pH 7.4是最佳binding
tech support的人也这么说
说buffer什么的不重要
重要的是pH
你的bead会不会饱和了？
然后我还遇到过的是除了融合蛋白以外还有27 kDa的GST-tag only表达
表达的还挺多
然后跟我的融合蛋白竞争
很容易就把柱子饱和了
哦，还有一个
你提到你的蛋白是inclusion body
我没用过N-lauroylsarcosine
如果好用的话回头汇报一下哈
不过我遇到inclusion body的时候用过Pierce的B-PerII
很好使
结果大部分都变成可溶了
当然我的那个例子比较极端
光用sonication是大多数都在pellet里
用B-PerII加sonication以后大多都在上清里
我一升的e.coli居然提了快100 ug的蛋白
% |
|
n***w
发帖数: 2405 | 32 前面已经发帖问了，有个前辈说there may be hydrophobic interaction between the
protein and the sepharose
beads so it's why it remained bound even after cleavage.
虽然说换载体换tag是个方法，我想知道市面上有没有除了sepharose的其他GST
binding matrix以减少hydrophobic
interaction.
我看了novagen和pierce的，他们的resin也和GE的sepharose差不多。。。
请问大家知道市面上还有别的不？
谢谢。 |
|
|
D******9
发帖数: 2665 | 34 你这个泡脚的结果现在不算了， 必须看REALTIME的fold enrichment. 除了normal IgG
外， 找一个跟你TF binding site 外 1-2 kb的地方做negative control， 再看你的
结果。 |
|
B*****e
发帖数: 1005 | 35 0.01%几乎与IgG的信号差不多了，这时候很难确定真实TF binding信号，还是背景，还怎么算fold enrichment？ |
|
z*******a
发帖数: 175 | 36 my comment on comments of Nemo and Biogene (see below). by the way, I like
these two names.
1. Once upon a time, I did not take any <0.03 % input as a real binding when
I used antisera not purified IgG.
But I saw some papers in top journals have data published like 0.00something
%input, really confusing. do
you two or 达人 have any comments?
2. I would say enrichment folds instead of fold enrichment. I should
calculate folds relative to normal serum
control when antisera are used or IgG control w... 阅读全帖 |
|
m******5
发帖数: 1383 | 37 Thank you much for your great contribution to my rookie question^^
I think non-related region negative control is convincing theoretically, if
not taking non- specific DNA precipitation into account: different DNA
region might have different tendencies of non specific precipitation (I also
learned that this issue can be simply avoid by using dynal beads)
A parable phenomenon is that when I was doing invitro protein interaction
assay such like GST pull-down and co-ip, the reviewer did not questi... 阅读全帖 |
|
a****o
发帖数: 1786 | 38 fold of enrichment is a better measurement than IP efficiency. fold greater
than 4 is a good number.
Low IP efficiency sometimes come from that YFP binds to certain target only
in a fraction of cells.
when
00something |
|
z*******a
发帖数: 175 | 39 I used solid tissue of animals. You are talking about binding of
exogenously transfected protein to
endogenous regulatory elements. I am wondering how you do transfection
efficiency control in the first place
if it is transient transfection. equal exogenously expressed protein makes
your data comparable between
treatments and repeats. I do NOT have confidence to get evenly transfected
efficiency all the time.
greater
only |
|
m******5
发帖数: 1383 | 40 I have another question: I didn't see much people have immune-precipitated
protein shown in their paper(even for those who did overexpression protein
immuneprecipitation) ,which is very very weird, because it should not be
very hard theoretically. without immune-precipitation efficiency, by which
they can convince the reviewer or themselves that the binding of DNA is
reasonable? |
|
f*********e
发帖数: 1144 | 41 终于有回复的了！！谢谢！
你做的competition assay时候是两个Ag同时一起加还是有先后顺序，而且wait for a
certian period of incubation time？Or it doesn't matter as the binding is
irrevisible? |
|
h**********r
发帖数: 671 | 42 Title: Immobilization of Lactococcus lactis to cellulosic material by
cellulose-binding domain of Cellvibrio japonicus
DOI: 10.1111/j.1365-2672.2010.04757.x
Journal of Applied Microbiology
Volume 109, Issue 4, pages 1274–1283, October 2010
My Email is m******[email protected]. Thank you so much! |
|
F*K
发帖数: 608 | 43 At this stage I want to know if it binds to DNA. |
|
c********r
发帖数: 189 | 44 1) FRAP to see the protein's mobility in nucleus.
2) cell fractionation to see if the protein only in DNA binding fraction |
|
|
c*t
发帖数: 1063 | 46 But the question asked is to confirm direct protein binding, I don't know if
I am "钻牛角尖"。 |
|
|
w***e
发帖数: 269 | 48 Your lists are fine. I don't think there is a "perfect" method. You just
have to cross-validate direct binding with results from multiple essays. |
|
c********r
发帖数: 189 | 49
Re
For direct binding, the only way is in vitro pull down assay with purified
proteins |
|
l****j
发帖数: 70 | 50 多谢提供的参考文献，
我研究的那个ER蛋白对线粒体有很多调节作用，比如ATP，O2，还有对线粒体复合体的
assembly都有影响，对钙离子也有调节作用
我想解决ER蛋白怎么对线粒体内膜有影响的，为甚么能binding在一起 |
|
