o****e
发帖数: 1011 | 1 我也是新手，顺便问一下，blocking solution里“一般”也要有tween吗？
（用“有tween的PBS/TBS”配,而不是用PBS/TBS配？）
Thanks。
~~~~倒是看到不少用0.05%的 |
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c*w
发帖数: 50 | 2 We know that when the Csurf. > cmc, the micelles form in the aqueous
surfactant solution.
My question is: take Tween 80 as an example. If the solution contains 90%
tween 80 and 10% water(v/v), is it still a micelle solution?
Thanks for the answer! |
|
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s*******n
发帖数: 10426 | 4 【 以下文字转载自 LeisureTime 讨论区 】
发信人: swordsman (买卖提的脊梁), 信区: LeisureTime
标 题: Marker Tween的过炸药文学奖吗？
发信站: BBS 未名空间站 (Thu Oct 11 15:39:32 2012, 美东)
马克吐温得过炸药文学奖吗？ |
|
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s******y
发帖数: 220 | 6 我是新手， 前人留下的protocol用的是0.1%, 有人推荐0.5%.
我们用pbs+tweeen， 这个浓度区别大吗？ tween到底是什么作用呢？
谢谢 |
|
s*********s
发帖数: 110 | 7 tween 20是detergent，用来洗脱unspecific binding
一般都用0.1% |
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s******s
发帖数: 13035 | 8 Tween是detergent，多用了背景低，但是可能信号也洗掉了
这个完全是更具antibody case by case，一般0.1%。
btw, 没人用PBS，都是用TBS的，PBS是直接做细胞用的 |
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s*********s
发帖数: 110 | 9 我们都是PBS+0.1%Tween 20,我所知道的lab大部分也都是PBS/T
偶尔有lab用TBS/T
或者有些cell signaling的抗体特别强调用TBS/T,如foxo的几个抗体 |
|
s*******n
发帖数: 10426 | 10 【 以下文字转载自 LeisureTime 讨论区 】
发信人: swordsman (买卖提的脊梁), 信区: LeisureTime
标 题: Marker Tween的过炸药文学奖吗？
发信站: BBS 未名空间站 (Thu Oct 11 15:39:32 2012, 美东)
马克吐温得过炸药文学奖吗？ |
|
w*******y
发帖数: 60932 | 11 Lord and Taylor is running a promo on Kids clothing, B1G1 50% off. Thought
the Columbia coats were noteworthy Some sizes might be ymmv with colors
selected.
Price will appear as $7.98 after you click on your color and size.
Second one adds to cart at 50% off.
Add coupon 'SAVE' to get prices down by another 15%.
Shipping is $5.95
COLUMBIA Girls 2-6x Full-Zip Fleece - Smart Value
$19.95 ($6.78 AC in cart for first one, $3.39 for second)
Link:
http://www.lordandtaylor.com/eng/Kids-kidsouterwear-Gir... 阅读全帖 |
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f***r
发帖数: 204 | 12 还在与PVDF战斗中，再次请教有经验的虾们：
终于看到了条带，但是巨多非特异带，比我要的带都强得多。在网上查到多家western系
统的trouble shooting，里面提到的好几条关于high background的建议让我大开眼界，
又困惑非常，因为矛盾很多：
1. 关于tween的使用：有一家建议blocking之后才用tween，否则tween会stick to the
membrane and cause high noise；但很多常规方法都没有提到这一点。
2. 关于blocking reagents和ab incubation buffer：有说dry milk能reduce
sensitivity，所以如果只用在二抗incubation就能很好降低noise，但也有很多全程用dr
y milk的，也有说dry milk与tween联合使用会提高noise的；有说BSA会造成高背景的，
也有说用BSA可以获得理想信噪比的……
3. 关于ponceau s染色：这次为确证transfer有效，用了ponceau s染色，但发现有资料
说ponceau s就算洗干净也会 |
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l*y
发帖数: 21010 | 13 FEATURES
SONGS
VIDEOS
LISTS
EMAIL
SHARE
TWEET
PIN IT
REDDIT
Neil Young: “Fuck You, Donald Trump”
Chris DeVille @chrisdeville | 4:59 pm
EMAIL
SHARE
TWEET
PIN IT
REDDIT
Neil Young
CREDIT: Ross Gilmore/Redferns
Neil Young, a fervent Bernie Sanders supporter, has not been fond of Donald
Trump using his song “Rockin’ In The Free World” at campaign events. Last
year Young asked Trump to stop using the song and issued a statement
explaining his opposition to the Republican candidate. That inspired Trum... 阅读全帖 |
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r****y
发帖数: 26819 | 14 说的是js啊
html5有针对event的声音和特效函数了吗？比如fade in， fade out，从某时刻开始
之类
动画来说，html5怎么做tween？比如motion的tween和shape的tween？ |
|
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i**e
发帖数: 19242 | 16 我感觉
tween的年龄开始“反抗”了
妈妈比较宠护有加，爸爸简单粗暴有加
1. 我希望自己的孩子11岁的时候，懂得什么事情是不可以做的；知道不要造成人为的
浪费，包括金钱，时间，精力和情绪；我觉着他懂得，不过人家不care，不原意自我规范
2. 我个人不认为是不小心的，我希望我的孩子在11岁的时候已经养成了垃圾顺手扔垃
圾桶的习惯
//说别人容易，我还在努力
3. 邻居里有tween/teen的娃，类似的“殃”大家都会时不时遭遭地；包括圣诞节的彩
灯，装饰，垃圾桶，草坪etc;也是抓不到什么证据地，叫孩子跟着大人一起收拾残局就
好了
4. 孩子有能力cleanup就让孩子做，没有就算了；没证没据的事
5. 这个是天性，谁急谁就是跟自己跟一家人过不去贝；要不你们商量，定个进餐的时
间好了
6. 不让打游戏是不是太过了？太简单粗暴了？这事宜疏不宜堵吧？
一家人好好坐下来谈谈，定些家庭规范？相互制约一下？ |
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P********L
发帖数: 31 | 17 以下是Pasadena图书馆主馆及分馆十二月部分活动预告. 所有活动免费对公众开放, 无
需出示借书卡或当地居民证明, 欢迎所有有兴趣的朋友前来参加. 部分活动因场地有限
须提前报名(此类活动会在预告中注明). 如有问题请致电(626)744-4066查询.
主馆和9个分馆地址: http://ww5.cityofpasadena.net/library/libraries/
更多活动请见: http://ww5.cityofpasadena.net/library/library-events/
--Family Events--
The Nightmare Before Christmas Fest
Create a new family tradition viewing The Nightmare Before Christmas. There
will be face painting, snacks, a photo op with a life-size Jack Skellington
and Zero, a movie facts “quiz show” with a... 阅读全帖 |
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l****e
发帖数: 359 | 18 你这个动画设计好没有？自己写好storyboard了吗？
同样的情节，有很多种方法可以解决，想烧钱的那就fbf，因为有很多动作
靠tween就是做不出，比如转身阿啥的。
想省钱，那就多定格，多用tween，但是效果自然就差很多。
还有一个办法，就是，你这个项目如果就是自己做着玩玩的，那就找人合作，
有不少人有技术没有好剧本，或者不介意和别人一起玩，到时候多署个名字
而已，有利益了一起分。
真要做动画片，code啥的用得很少很少，但是需要好的美工和配乐。而且如
果不想一个项目作3年，那就需要多个人一起做，需要找一个已经有配合基础
的工作室。 |
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r****o
发帖数: 3918 | 19 特效估计要用after effects做。
做两分钟的tween也挺耗时的实话说。。
那些小天使飞来飞去，需要花俏的，那一个画面要好几个tween呢
尤其是这个东西没啥艺术性，还放不进portfolio。volunteer不久图一个自由创作能放
作品集里面嘛。。
不过我在水木的arts版看见过有人求做这个
最后好像有人接了，价钱大概1k这样的吧。
楼主可以去试试 |
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r****o
发帖数: 3918 | 20 特效估计要用after effects做。
做两分钟的tween也挺耗时的实话说。。
那些小天使飞来飞去，需要花俏的，那一个画面要好几个tween呢
尤其是这个东西没啥艺术性，还放不进portfolio。volunteer不久图一个自由创作能放
作品集里面嘛。。
不过我在水木的arts版看见过有人求做这个
最后好像有人接了，价钱大概1k这样的吧。
楼主可以去试试 |
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a**p
发帖数: 49 | 21 30 micron meter
well
u can do it in floating sections for staining
with triton rather than tween
staining overnight at 4 degree
TWEEN is too weak
if background too high
reduce concentration of 2nd Ab as well
not necessarily the 1st |
|
s******y
发帖数: 28562 | 22 洗的时候肯定要用tween, 不然就洗不干净了。
但是加抗体的时候不应该加tween, 不然有些比较弱的抗体的结合会受到影响。
我不知道你们做的是什么蛋白，但是你们的红色的带怎么这么多啊？
要么就是你们用的一抗太差，要么就是非特异结合（背景信号的一种）了。 |
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s********n
发帖数: 248 | 23 M-CSF培养的bone marrow macrophage，stimulate以后western blot 磷酸化的AKT/ERK
/STAT1，其他两个都blot的很好，但是pAKT总是没有想要的带，貌似出来的带都是二抗
结合的非特异性带。
请问版上高人，有blot过pAKT的没有？能否帮我看看问题出在什么地方了啊？
我用的WASHING BUFFER是TBST with 0.05%Tween 20，
BLOCKING BUFFER:5%BSA/TBST,
一抗就dilute在blocking buffer里边。
protocol:
室温block 1h，一抗4度过夜，洗3次×10min；二抗室温30min，洗3次×15min。
我初步怀疑是tween 20量加多了，又看到班上有用pbs做washing buffer的，貌似pbs更
温和一些，不知道用pbs会不会好一些？
小女子多谢了先！ |
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a******y
发帖数: 63 | 24 Less than 10% of DMSO should be fine for injection. BTW, 20% of Tween 80 or
Tween 20 may also be included. |
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s******y
发帖数: 28562 | 25 http://www.piercenet.com/browse.cfm?fldID=02020104
Substances compatible with the Thermo Scientific Coomassie Plus Protein
Assay. The following is a short list of compatible substances and their
concentrations that the BCA Protein Assay can tolerate. For a more complete
list please refer to the instruction booklet for the product or view our
Table of Compatible Substances for a comparison of substance compatibilities
among several different protein assay methods.
Substance Compatible Conc. ... 阅读全帖 |
|
l*******n
发帖数: 1867 | 26 Post a comment
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Thanks for posting. Would you like to edit your profile?
straso10 french frogs
0 minutes ago | Like | Report abuse
SimpleStupid Was DSK visiting at the time? Probably has a severe case of DSB
by now, which could result in an explosion.
2 minutes ago | Like | Report abuse
gulfstream69 God hates the French too.
3 minutes ago | Like | Report abuse
billfusion For inventing the manage ah tius? (3some)
0 minutes ago | Like | Report abuse
Madukali Proke, ... 阅读全帖 |
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f**x
发帖数: 4325 | 27 most popular tween-engine plane producer.
aircraft is gonna be Da Bai Cai soon |
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l****z
发帖数: 29846 | 28 by Matthew Boyle 25 Mar 2013, 8:00 AM PDT
Sasha and Malia Obama are quietly vacationing at the Atlantis resort on
Paradise Island in the Bahamas, Breitbart News has learned.
A source tipped Breitbart News off to the First Daughters’ spring vacation,
which was not publicly announced or reported.
Breitbart subsequently confirmed President Barack Obama’s daughters’ trip
with other sources. Both the White House and the Atlantis resort declined to
confirm the report or comment, but another guest prov... 阅读全帖 |
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x*******5
发帖数: 1335 | 29 Titles in this series are assigned to academically qualified individuals who
engage in teaching, research or other creative work, and University and pub
lic service to the same extent and at the same level of performance as those
holding corresponding titles in the Professor series in the same department
. Such assignments, however, shall be made only under conditions and restric
tions (see
APM - 270-16, 270-17, and 270-20) which serve to make a clear distinction be
tween appointments in this se... 阅读全帖 |
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C********n
发帖数: 6682 | 30 给你来个反面的
http://healthland.time.com/2010/12/27/study-earphone-loving-teens-can-hear-just-fine/
Study: Earphone-Loving Teens Can Hear Just Fine
Teens aren't always the best listeners but a new study shows that their
hearing may not be to blame.
Despite the ubiquity of ear- and headphones for listening to music or cell
phone conversations, a survey of more than 4300 12 to 19 year olds found no
significant increase in hearing loss between 1988-1994 and 2005-2006. The
study, which questioned adolesce... 阅读全帖 |
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a***s
发帖数: 616 | 31 之前有人问同时开personal和business卡能否拿到2*35,000 = 70,000 miles。
申请链接页面给的Terms and Conditions中说：
This offer is valid for first-time Delta SkyMiles Cardmembers only. If you a
lready have or had any other Delta SkyMiles Credit Card from American Expres
s Account(s), you are eligible to receive only the incremental difference be
tween the Welcome bonus miles offered herein and the amount offered with suc
h other Account(s).
也就是说可能第二张卡的bonus是25,000 miles而不是35,000 miles。
根据以往经验，biz卡和personal卡属于不同类型的卡，bonus并不冲突 |
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p****x
发帖数: 4260 | 32 how much is your score now? My credit socres from Creditkarma always fall be
tween 720-760, however, my real score is much more (>800) :D
anyway, if your score is more than 740, I don't see any reason why you shoul
d worry about that. |
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w********t
发帖数: 18315 | 33 1, They hardly remember anything before age 2.
2, Your toddler is too small to understand what u called the @#$#
who cut you off on the parkway.
3, Your tween is big enough to understand what u said but smart
enouth not to repeat it in front of grandma.
4, Sooner or later ur child will come to see a large purple dinosaur
as unsuitable marriage material.
5, There must be at least some trace minerals in chips.
6, Losing the spelling bee or suffering through a few years of
braces actually does buil |
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m**y
发帖数: 18546 | 34 老大到了tween的年龄，天天和我顶嘴。今天还质问我当初为什么要把她生下来。天地
良心，怀她的时候我才22岁，定着
多少反对和压力才把她生下来的啊！晚上老二又和我闹绝食，什么都不肯吃。我左骗右
骗手都摇断了也不肯开口。最后我受
不了了，冲着苍天大吼了一声来发泄内心的郁闷。吼完我就后悔了。哎！我这妈怎么当
的。幸好没有吓到宝宝。 |
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b*******y
发帖数: 12791 | 35 我买了个减价的ju ju be be tween。比较小，分层挺多，很方便，反正我觉得暂时够
用了。 |
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a*****g
发帖数: 19398 | 36 According to Common Sense Media, tweens log 4 1/2 hours of screen time a day
, seven days a week, 52 weeks a year. For teens, it's even higher: nearly
seven hours a day. And that doesn't include time spent using devices for
school or in school.
From babies with iPads to Chromebooks in classrooms, digital devices seem
more ubiquitous every year. And one of the hottest issues today in both
parenting and education circles is the proper role of electronic media in
children's lives.
There's research ... 阅读全帖 |
|
m****o
发帖数: 837 | 37 Google出来的：
how often should kids bathe? Once every day or two, unless your child is
particularly active or messy is enough, says Christopher Tolcher, MD,
clinical assistant professor of pediatrics at the University of Southern
California School of Medicine. “Bathing daily becomes more important for
tweens and teens,” he adds, “because issues with changes in the skin and
body odor become more evident.” |
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i**e
发帖数: 19242 | 38 欧昨天很有辛地近距离接触了一位tween
感受了一哈兹
i don't care, who cares, i don't know, i don't want to be with you
之态度和气场
//欧一哈兹没有了自己的娃到时候不会这样的气势 |
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i**e
发帖数: 19242 | 39 有辙啊！
不过呢，你
一无权，二无责任，三无权威，四无感情，五无经验
你就有气愤21 :)
欧不晓得怎么帮你使劲，除了把自己的日子过好，学习跟她相处（这样还有可能她听点
你的）
虽说，欧也不得不混得长辈希希地，越来越觉着，很多时候尊重是自己挣来的(earned)
tween，就是9岁之后13岁之前的娃
从小慢慢要求，慢慢教育出来的 :)
么？ |
|
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m**k
发帖数: 18660 | 41 展开说说tween and teen的督导?
如何对待游戏，运动。学业.课外。竞赛。。友情? |
|
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s***l
发帖数: 2236 | 43 ☆─────────────────────────────────────☆
qingqingfeng (qingqingfeng) 于 (Fri Jan 18 15:39:56 2013, 美东) 提到:
儿子11了，4岁半来美，现在初中一年，去年8月刚搬家到新的学区。在新学校总是出现
问题，以前小学不这样的。刚开始不会打开locker之类的事，就是手忙脚乱也过来了。
有时会为了小事，他就急。
最大的问题是说话不合时宜，经常说出让人意外话。一次，他说他的钢琴老师，你
smell like a Burger，我立刻打住他，可是已经说了，我很尴尬，后来老师就不在教
他弹琴了。我们已经发现他这个问题，总是在教他，可是，这好像是天生的，他改不了
。在新学校，一次，他见几个孩子在闲聊，他问在旁边的老师，他们在说什么，老师忙
，就说not your business,他就很生气，追上老师说，你为什么这么rude，老师当然不
会高兴，把他教育了一顿。另一次，他在课上举手想回答问题，第二个问题，又举手，
最后老师叫他了，结果他站起来问老师问什么才叫他，老师就发飚了，儿子就哭了。
... 阅读全帖 |
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a*****g
发帖数: 19398 | 44 According to Common Sense Media, tweens log 4 1/2 hours of screen time a day
, seven days a week, 52 weeks a year. For teens, it's even higher: nearly
seven hours a day. And that doesn't include time spent using devices for
school or in school.
From babies with iPads to Chromebooks in classrooms, digital devices seem
more ubiquitous every year. And one of the hottest issues today in both
parenting and education circles is the proper role of electronic media in
children's lives.
There's research ... 阅读全帖 |
|
|
l*******o
发帖数: 12469 | 46 Posted By crranky ~ 21st August 2012
Back in 2007 I started a free Blogspot web site in the gaming space. Within
6 months, via Google Adsense (and a couple other rev sources) it was bring
in $3,800 a month. (CORRECTION: the 3,800 was not Google Adsense alone).
A year later the site was receiving 110,000 uniques, 1.5 million page views
and over $10,000 a month (with multiple rev sources). This juggernaut was a
simple blog of mostly daily posts about a certain game and in most cases
those blog pos... 阅读全帖 |
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w****q
发帖数: 134 | 47 月底离开芝加哥，低价出让所有家具：沙发，QUEEN SIZE床(ROOM PLACE)，TWEEN SIZE
床，电视机，餐桌和椅子，空调(GREE 8000BTU)，微波炉，电扇，加湿器，净水器，全
自动热水壶；另有女式单车（自行车），所有的东西都很新，有意者站内联系。 |
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w****q
发帖数: 134 | 48 月底离开芝加哥，低价出让所有家当：沙发，TWEEN SIZE床，电视机，餐桌和椅子，空
调(GREE 8000BTU)，加湿器，净水器，全自动热水壶；另有女式单车，有意者站内联系。 |
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y*z
发帖数: 2555 | 50 By Bessel Kok
Over the last few days, I have been flooded with telephone calls and ele
ctronic messages asking me to comment on the recently announced match be
tween Veselin Topalov and Vladimir Kramnik. I was aware that Joel Lautie
r had been working on this and I must thank him and anybody else who wor
ked hard for this reunification match.
My initial reaction was of great joy for the players as well as for the
whole chess world! At long last the possibility exists for a match to be
played, th |
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