n***w
发帖数: 2405 | 1 最近用GST beads，有个疑问，
bacteria pellet我没有用PBS dissolve，而是用了自己配的一个solution, 内含1.5%
N-lauroylsarcosine，25mM TEA和
1mM EDTA (pH 8.0)。
最近发现binding有些问题，因为ideally, binding前后的supernatant应该最明显的区
别就是target protein少了，这个我
没有观察到。
我在想到底是哪个地方出了问题。
有经验的同学指点一下～谢谢！ |
|
n***w
发帖数: 2405 | 2 嗯，有可能饱和了。
我昨天把1st binding后的supernatant又加beads了，看好没好些今天。
我觉得N-lau还是挺好使的。能solubilize大部分pellet。
我500ml的culture提了差不多160ug的蛋白。。。
我下次买点你说的B-Per试试好了～ （不过看成分也差不多）
我现在的protocol基本上是：(For 500ml culture)
1. thaw pellet on ice
2. add 18ml GST buffer (配方见上)，add 1% PI, add 2ml lysozyme (10mg/ml)，
mix on ice for 30 seconds
3. pass thru 20G syringe to reduce viscosity, also add DNAse 40ul (1mg/ml).
4. sonication (it will become clear)
5. 10000rpm, 40min，4deg
6. collect supernatant for GST beads binding
不过我这次还是将G... 阅读全帖 |
|
f*********e
发帖数: 1144 | 3 我想请教一下版上有Ab Ag binding经验的如下问题：
我的样品（Ag)里面有不同的species 都可以跟Ab 作用，因为它们都有相同的epitope.
比如，A, B,C,D 都可以跟Ab作用。但是[A]>[B]>[C]>[D]. 不知道到底浓度差别多大
，估计最大最小之间有20倍。
我先在的问题是，after incubation for two hours, A可以被capture, B也可以，但
是C和D都没有。所以我就在想是不是Ab的位点可能都被饱和了？有没有什么方法可以证
明确实是因为这个原因？比如after incubating with the real sample, 然后加入更
高纯度的已知Ag(E)with better affinity，看会不会再有这个E的出现？但是首先需要
确认的是这个binding是不是reversible,那个E会不会kick off previously captured
A and B?
多谢多谢！！！！！
|
|
l****j
发帖数: 70 | 4 我从细胞中分离纯化的ER和mitochondria，检测过marker蛋白没有相互contamination，
细胞转染过 HA-tag 的 ER protein，WB验证只在ER和MAM(mitochondria associated
ER membrane)存在，纯化的线粒体中没有发现此蛋白。
在whole cell lysate用 HA-agarose Immunoprecipitation,在同一张膜上
immunoblotting 两个 mitochondria inner membrane protein（mito protein 1，2）
竟然发现和mito protein 1 有 binding,
分别在不同细胞器中的蛋白质，有可能binding吗？ |
|
c***y
发帖数: 615 | 5 最近比较6个样本的老鼠RNAseq数据，有些基因 (200 左右)，和17个transcription
factor 表达高度相关 （pearson correlation >0.98）。 但是，各类transcription
factor binding 数据库都无法表明any of those transcription factors has
binding site a the promoter of those 200 genes. 该如何解释这种情况呢？
多谢了！！ |
|
b******n
发帖数: 23 | 6 I need to coat polystyrene plastics with gamma-
methacryloxypropyltrimethoxysilane(bind-silane) without breaking the double
bond in Bind-silane. Don't know how and wonder if anyone can help. many
thanks! |
|
w*******y
发帖数: 60932 | 7 CD / DVD Sleeves w/ 2 Binding Holes, 100pcs, Assorted Colors $1.99. Free
shipping from EveryDay Source. ( Starts at 11:30 AM Eastern )
Starts 11:30 AM Eastern
$7.99 - $6 off code = $1.99
Use Promo Code S0613PPKT
CD / DVD Sleeves w/ 2 Binding Holes, 100pcs, Assorted Colors:
http://www.everydaysource.com/product/DD/80043?esaffcojutxt1106 CJ 80043 07
|
|
t****x
发帖数: 9441 | 8 为毛不是雪主red body nude body five flower big bind |
|
w*****r
发帖数: 2540 | 9 这个卖家没说是那种binding，同一个型号有两种，一种是适合旧板子，一种是适合新
板子。他也没有
说明是那种，图片也看不出来，就说跟官网一样的配置,而且全新。然后我就拍了一个
（下的bid).东西
拿到之后发现是适合旧板子的（我的是新板子）， 而且上买你有个地方有特别明显的
擦痕，面积还不
小。昨天给卖家回信，现在还没回。我还有什么办法呢？直接找ebay要求refund? 请指
教，谢谢 |
|
d*******l
发帖数: 2567 | 10 【 以下文字转载自 NanoST 讨论区 】
发信人: demonbull (牛魔王/Darth Vader), 信区: NanoST
标 题: 包子酬谢：谁有纳米碳管的计算DOS的tight-binding模型
发信站: BBS 未名空间站 (Tue Jun 23 17:01:15 2009, 美东)
单壁，多壁都行，最好有多壁的（>2就行）。
有C,Matlab程序最好了。 |
|
|
S**Y
发帖数: 136 | 12 I don't quite understand.
when i am reading os textbook, it says the DOS bind address
at compile time. So when a DOS excecutable file is copy/pasted to another ma
chine, it can not run because the address might not be available at the new
machine.
Who can explain this? Really confused...
Thanks! |
|
n******5
发帖数: 91 | 13 谢谢
我主要关心的是 Apr 30这个截至日期，是否需要拿到bank approval才算binding
contract? |
|
g*****g
发帖数: 3623 | 14 卖方接受offer，双方签字，然后inpsection and get mortagage?
or
final closing?
tax credit now applies to sales occurring on or after January 1, 2009 and on or before April 30, 2010. However, in cases where a binding sales contract is signed by April 30, 2010, a home purchase completed by June 30, 2010 will qualify. |
|
p****n
发帖数: 149 | 15 I do hope I can get the first-time homebuyer credit. Is it possible to file
a written binding contract in two days? |
|
n****e
发帖数: 1403 | 16 一般大家签合同的时候，从binding agreement date到closing date大概多少天？ |
|
m*****j
发帖数: 499 | 17 我们正在买一房子，下offer，卖家counter offer，我们又counter offer回去，但是在
我们的counter offer过期时间之前没有收到任何消息。本来以为没戏了，但是忽然age
nt发信说要inspection。
我们很不爽，房子也不想要了，agent肯定要fire了。但是不知道这种情况（估计卖家接
受了，但是我们没得到消息）算不算binding，我们现在可以说不要了吗？
地点在OR。多谢各位指教！ |
|
m***5
发帖数: 83 | 18 www.utmb.edu/international/visainfo/I485checklist.pdf
A good checklist for binding.
Cheers. |
|
g********e
发帖数: 1638 | 19 你把支票放到第一页，其实就是左上角，下边放着E-file的第一页吗，
或者你用一个很小的clip把check单独bind到E-file的第一页。
第二页是check-list
我的Eb1A pp-ed 140+485 都ok了
另外尽量一起递交485. 一起捆绑拿到很快的。尽量不要单独递交140.
因为485需要的材料很少。就是出生公证和体检需要准备。
出生公证找家里人办理，很快的，就几天，和体检需要的时间差不多。
其他的都是表格什么的。 |
|
g********e
发帖数: 1638 | 20 那个Efile产生的pdf文件的第一页有个条形码
这个是要放到最前的，可以让pp小组马上知道你是efile得。
然后你支票用个小的binder或者直接把支票放到有条形码的最左上方，
用大夹子一起bind
那个checklist最好有，方便门房的人看看你提供的资料够不够，另外你自己在邮局邮
寄的最后时刻也可以检查一下有没有东西忘了放进去。
另外千万要注意
（1） 该签字的地方一定要签字，
（2）支票上的钱 amount是对的
这两个是最重要的，在邮局做最后check。 |
|
G*******1
发帖数: 137 | 21 审稿机会：-Retinol binding protein-4 (RBP4) levels 在肥胖儿童和以及与脂肪肝
的sonographic grading关系
有意向者请站内联系。谢谢 |
|
w****u
发帖数: 1078 | 22 Where can I get the things for binding after tow holes are punched?
The thread-like stuff.
Thanks. Baozi for the first one with the right answer. |
|
h****a
发帖数: 628 | 23 那位次有NIW binding 模板?
装订的时候, 有什么要注意的吗? 一定要index吗?
xiexie |
|
r*******t
发帖数: 8550 | 24 中国人的地产商喜欢叫人出$5000-$10000
美国人的地产商一般叫人出$500-$1000
买家出多binding offer钱的话，对卖家和经纪都稍微有利，有钱赚大家分。 |
|
k******p
发帖数: 96 | 25 Ski rack: Dynamic One Flexcap 160cm
Ski binding: Tyrolia SL110
• Stand height 21 mm
• DIN 3-11
• ABS – ANTI BLOCKING SYSTEM
• SL Heel
• SL Toe with TRP System
• FULL DIAGONAL
• Dura-Coating
ELIM 125 ski poles
All in good condition. Ask for $200.00
Please email me if you are interested. |
|
J******h
发帖数: 6102 | 26 hiking到山顶，然后滑雪下来。但是不愿意多背一双滑雪鞋。
请问有登山鞋和ski的binding吗？ |
|
|
J******h
发帖数: 6102 | 28 binding with mountaineering boots, leather or plastic |
|
N***G
发帖数: 408 | 29 rei, sports authority, online, local stores.
我带过snowboard with bindings装在一般的snowboard bag里. 算一件行李，传送带上
没等到，结果到大件处领出来的，没有太大问题。 |
|
n*******3
发帖数: 313 | 30 【 以下文字转载自 ChuanYu 讨论区 】
发信人: nancy0123 (又见rainbow), 信区: ChuanYu
标 题: I have a pair of skis with bindings to give away, anybody interested?
发信站: BBS 未名空间站 (Thu Sep 24 16:15:04 2009, 美东)
don't want to bother taking with me into the new apartment. Anyone
interested can contact me.
BTW, It's for men's skis, 180mm. |
|
T********e
发帖数: 8631 | 31 如果需要jig and drill的话没办法，一共就几个螺丝干嘛要花那个冤枉钱。binding
mounting还有好多讲究，如果你不自己装以后也不会调的。
另外还在选购tuning kit，wax，etc，碰到高手也可以顺便聊聊。 |
|
T********e
发帖数: 8631 | 32 I understand some skis require special tools to mount, but in my case, there
's only one position you can put the plates on with pre-drilled hoes on the
mounting platform. The toe and heel pieces are ridged and marked for each
shoe size to the millimeter. The only thing you can change is center/
forward mounting position, forward pressure and toe height adjustment and
DIN setting. All those you are supposed to experiment and adjust as time
goes by. And we all know how binding release and you |
|
m****g
发帖数: 3975 | 33 NE的人工费真贵, 我上一副SKI还是在OREGON做BINDING的, 价钱只有一半左右, 特价
SPRING PASS 才$80, 雪况还要好. COLORADO的DEAL也比较多.
要滑雪还得去西部, 本地就用高山和越野滑树林就成了. MOGUL这里的很ICY. |
|
d****r
发帖数: 300 | 34 I just installed bindings for my new ski. It is an integrated system. So it
is easy. By following the instruction, the installation is very
straigtforward and toolless. The last step is to adjust the din number.
Google "ski din", you will get some charts to get your correct din number. I
don't know how a professional guy can do more to make the installation "
professional" :-) |
|
|
|
n*****a
发帖数: 1327 | 37 a compare of forward pressure indicator with and without boot in.
you should adjust the forward and rear pieces of the binding so that when
you insert the boot, some marker or line shows up near the middle of the
indicate window.
driver |
|
B*******a
发帖数: 410 | 38 Thank you. For my binding there is only boot size indicator. I cant find any
mark regarding to sole length. |
|
|
v****e
发帖数: 19471 | 40 thanks, bro. I just got the K2 -- strong brand, easy to sell if doesn't work
for me. regarding the bindings, I'll go to the loacal shop to have them ins
talled and tuned -- my ld is having her skis tuned there, anyways.
good |
|
v****e
发帖数: 19471 | 41 I got the K2 Apache 167CM with the Marker M11 TC bindings. Looks like a good
deal for a set of all-mountain gear for intermediate. If they don't work fo
r me (which is unlikely considering I really can't tell what is working and
what isn't), the resell value shouldn't be too bad. I can't complain about s
pending a total of $300 on this $1000 set.
both
skis, |
|
v****e
发帖数: 19471 | 42 那太好了，看来能管几年。俺整齐了全套：
Boots: Nordica Supercharger Ignition
Ski: K2 Apache Stryker 167cm
Bindings: Marker M 11 TC Piston
税后总成本：$750
diamond
good
from |
|
R******e
发帖数: 2254 | 43 Nice, I spent about $600 on my boots, skis and binddings
I used my boots for more than 3 seasons and used my skis for more than 2
seasons
I do not intend to update them, at least for the next couple of years.
Actually, maybe I do not buy any Alpine skis any more, I am switching to
Randonee skis. |
|
r*******1
发帖数: 4394 | 44 if it come with intergrated system (binding rail has been installed on skis)
, then you can install by yourself within 10 mins. Check your DIN setting,
and adjust accordingly.
Still recommend do it by pro ($40). |
|
k**t
发帖数: 1968 | 45 If the skis came with an integrated bindings package, you can mount them
yourself by reading instructions. Otherwise, you have to take them to a ski
shop. Search your local yellow book to find one. |
|
y*******o
发帖数: 6632 | 46 I bought two boots but I do not know what kind of bindings I should choose
to fit my boots. Any one can give me some advices? Thanks a lot for your
help. |
|
j******e
发帖数: 1424 | 47 Binding match boots. check your boot's sole size |
|
k**t
发帖数: 1968 | 48 That's ok. As long as the bindings can be set to your din #. |
|
d***e
发帖数: 192 | 49 据说burton的binding只有3个孔可以固定。是不是这样的话就不方便固定在非burton的
雪板上了？ |
|
