concentration and kinds of detergent for cell (293T) lysis?# Biology - 生物学
j*g
1 楼
Hotel的信用卡哪张比较好？ 能给个介绍或者链接什么的么
H*i
2 楼
http://www.immigration-law.com/Canada.html
Senator Grassley lifted hold on this bill removing the first hurdle. However
, the bill has yet to face two challenges in the Senate, not to mention any
other potential problems in the House. One is potential troublesome or
controversial amendments that can be introduced on the floor including the
Senator Grassley's amendment. The second is potential additional objections
to the bill on the floor and potential problem of passing motion to cloture
by at least 60 votes. For the first problem, it has been made clear that the
Irish E-3 will no longer be a threat as it appears that this was an
agreement of Sentor Schumer with the Irish community and probably part of
his agreement with Senator Grassley. However, nothing can be ruled out at
this point as to what other controversial amendments can be introduced and
passed as part of the bill which may not be well swallowed by the Republican
House. As for the second problem, AILA has reported that there are some
Senators who may object to the bill on the floor, forcing motion to cloture
requiring 60 votes. No names of the Senators are known at this point but it
appears it is a credible information. On top of all of the foregoing hurdles
, clock keeps ticking with the remaining Congressional sessions running out
quickly. Supporters of the bill hope to see this bill passed before the
election in November, but time poses a huge threat to such wishful thinking.
If it fails to pass before the election, it should be made through the lame
duck session during the nation's most celebrated holiday season. Otherwise,
the bill will die at the end of the year since the 112th Congress will be
closed and all the pending legislative bills will die along with the sunset
of the 112th Congress. Who knows, though? A miracle can happen.
Senator Grassley lifted hold on this bill removing the first hurdle. However
, the bill has yet to face two challenges in the Senate, not to mention any
other potential problems in the House. One is potential troublesome or
controversial amendments that can be introduced on the floor including the
Senator Grassley's amendment. The second is potential additional objections
to the bill on the floor and potential problem of passing motion to cloture
by at least 60 votes. For the first problem, it has been made clear that the
Irish E-3 will no longer be a threat as it appears that this was an
agreement of Sentor Schumer with the Irish community and probably part of
his agreement with Senator Grassley. However, nothing can be ruled out at
this point as to what other controversial amendments can be introduced and
passed as part of the bill which may not be well swallowed by the Republican
House. As for the second problem, AILA has reported that there are some
Senators who may object to the bill on the floor, forcing motion to cloture
requiring 60 votes. No names of the Senators are known at this point but it
appears it is a credible information. On top of all of the foregoing hurdles
, clock keeps ticking with the remaining Congressional sessions running out
quickly. Supporters of the bill hope to see this bill passed before the
election in November, but time poses a huge threat to such wishful thinking.
If it fails to pass before the election, it should be made through the lame
duck session during the nation's most celebrated holiday season. Otherwise,
the bill will die at the end of the year since the 112th Congress will be
closed and all the pending legislative bills will die along with the sunset
of the 112th Congress. Who knows, though? A miracle can happen.
b*y
4 楼
I am over-expressing GFP-tagged protein in 293T cell and planning to IP
using anti-GFP antibody to see what interaction partners are pulled down.
Anyone can suggest the appropriate detergents and their concentrations for
breaking the cell while not interfering with protein-protein interactions?
Thanks a lot!
using anti-GFP antibody to see what interaction partners are pulled down.
Anyone can suggest the appropriate detergents and their concentrations for
breaking the cell while not interfering with protein-protein interactions?
Thanks a lot!
r*c
5 楼
spg
【在 j**********g 的大作中提到】
: Hotel的信用卡哪张比较好？ 能给个介绍或者链接什么的么
【在 j**********g 的大作中提到】
: Hotel的信用卡哪张比较好？ 能给个介绍或者链接什么的么
c*l
6 楼
Thanks for the info.
Let's cross our fingers.
Let's cross our fingers.
t*e
7 楼
还是太贵了
s*s
8 楼
这个多了，取决于你作用强不强，背景怎么样了。没具体做实验没人知道，你从弱的一
点点往上做吧。
【在 b******y 的大作中提到】
: I am over-expressing GFP-tagged protein in 293T cell and planning to IP
: using anti-GFP antibody to see what interaction partners are pulled down.
: Anyone can suggest the appropriate detergents and their concentrations for
: breaking the cell while not interfering with protein-protein interactions?
: Thanks a lot!
点点往上做吧。
【在 b******y 的大作中提到】
: I am over-expressing GFP-tagged protein in 293T cell and planning to IP
: using anti-GFP antibody to see what interaction partners are pulled down.
: Anyone can suggest the appropriate detergents and their concentrations for
: breaking the cell while not interfering with protein-protein interactions?
: Thanks a lot!
g*i
9 楼
Re
★ 发自iPhone App: ChineseWeb - 中文网站浏览器
【在 r****c 的大作中提到】
: spg
★ 发自iPhone App: ChineseWeb - 中文网站浏览器
【在 r****c 的大作中提到】
: spg
p*j
10 楼
yes, cross our fingers.
h*2
11 楼
疯狂
花20-30块找个430-500w corsair或者ocz或者enermax，挺10年
花20-30块找个430-500w corsair或者ocz或者enermax，挺10年
l*1
12 楼
CA630 1%
Immunoprecipitation of IMC components. Highly synchronized late-stage
parasites were purified over 70% (wt/vol) Percoll, and proteins were
extracted by
incubating them on ice for 15 min in RIPA buffer (1%
octylphenoxypolyethoxyethanol
[IGEPAL CA-630], 150 mM NaCl, 0.5% sodium deoxycholate
[DOC], 0.1% sodium dodecyl sulfate [SDS], 50 mM Tris, pH 7.4). Soluble and
insoluble fractions were separated by centrifugation at 12,000 g for 10
min at
4°C. Soluble lysates were incubated for 1 h at 4°C using the rabbit
antibodies
anti-PfGAP45, anti-GFP, and anti-MTIP and preimmune serum. A 1:1 ratio of
protein-A/G–Sepharose was used for immunoprecipitation (IP). IP lysates
were
separated by SDS-PAGE and analyzed by Western blotting using mouse
antibodies
against GFP, PfGAP45, or PfMTIP. Western blots were developed as
described above.
Eukaryot Cell. 10: 556-64.
Yeoman JA et al.
Tracking Glideosome-associated protein 50 reveals the development and
organization of the inner membrane complex of
Plasmodium falciparum. (2011).
http://www.ncbi.nlm.nih.gov/pubmed/21239623
【在 b******y 的大作中提到】
: I am over-expressing GFP-tagged protein in 293T cell and planning to IP
: using anti-GFP antibody to see what interaction partners are pulled down.
: Anyone can suggest the appropriate detergents and their concentrations for
: breaking the cell while not interfering with protein-protein interactions?
: Thanks a lot!
Immunoprecipitation of IMC components. Highly synchronized late-stage
parasites were purified over 70% (wt/vol) Percoll, and proteins were
extracted by
incubating them on ice for 15 min in RIPA buffer (1%
octylphenoxypolyethoxyethanol
[IGEPAL CA-630], 150 mM NaCl, 0.5% sodium deoxycholate
[DOC], 0.1% sodium dodecyl sulfate [SDS], 50 mM Tris, pH 7.4). Soluble and
insoluble fractions were separated by centrifugation at 12,000 g for 10
min at
4°C. Soluble lysates were incubated for 1 h at 4°C using the rabbit
antibodies
anti-PfGAP45, anti-GFP, and anti-MTIP and preimmune serum. A 1:1 ratio of
protein-A/G–Sepharose was used for immunoprecipitation (IP). IP lysates
were
separated by SDS-PAGE and analyzed by Western blotting using mouse
antibodies
against GFP, PfGAP45, or PfMTIP. Western blots were developed as
described above.
Eukaryot Cell. 10: 556-64.
Yeoman JA et al.
Tracking Glideosome-associated protein 50 reveals the development and
organization of the inner membrane complex of
Plasmodium falciparum. (2011).
http://www.ncbi.nlm.nih.gov/pubmed/21239623
【在 b******y 的大作中提到】
: I am over-expressing GFP-tagged protein in 293T cell and planning to IP
: using anti-GFP antibody to see what interaction partners are pulled down.
: Anyone can suggest the appropriate detergents and their concentrations for
: breaking the cell while not interfering with protein-protein interactions?
: Thanks a lot!
n*y
13 楼
不容易呀！Let's bless each other.
However
any
objections
cloture
the
【在 H******i 的大作中提到】
: http://www.immigration-law.com/Canada.html
: Senator Grassley lifted hold on this bill removing the first hurdle. However
: , the bill has yet to face two challenges in the Senate, not to mention any
: other potential problems in the House. One is potential troublesome or
: controversial amendments that can be introduced on the floor including the
: Senator Grassley's amendment. The second is potential additional objections
: to the bill on the floor and potential problem of passing motion to cloture
: by at least 60 votes. For the first problem, it has been made clear that the
: Irish E-3 will no longer be a threat as it appears that this was an
: agreement of Sentor Schumer with the Irish community and probably part of
However
any
objections
cloture
the
【在 H******i 的大作中提到】
: http://www.immigration-law.com/Canada.html
: Senator Grassley lifted hold on this bill removing the first hurdle. However
: , the bill has yet to face two challenges in the Senate, not to mention any
: other potential problems in the House. One is potential troublesome or
: controversial amendments that can be introduced on the floor including the
: Senator Grassley's amendment. The second is potential additional objections
: to the bill on the floor and potential problem of passing motion to cloture
: by at least 60 votes. For the first problem, it has been made clear that the
: Irish E-3 will no longer be a threat as it appears that this was an
: agreement of Sentor Schumer with the Irish community and probably part of
a*o
14 楼
要是9.99 AR哥就买叁，呵呵。
【在 J*********n 的大作中提到】
: http://www.newegg.com/Product/Product.aspx?Item=N82E16817151120
【在 J*********n 的大作中提到】
: http://www.newegg.com/Product/Product.aspx?Item=N82E16817151120
l*1
15 楼
要学Yamanaka Lab PD Takahashi iPS 4 因子 筛选那样
样品攒足了 分成 50份
先高浓度 IP down 整死那个 No. 50
再一步一步 浓度 减法 Immunoprecipitate 下一份 No.49
>No.48
.
.
.
>
>
>
>No. 01
也未尝不可
【在 s******s 的大作中提到】
: 这个多了，取决于你作用强不强，背景怎么样了。没具体做实验没人知道，你从弱的一
: 点点往上做吧。
样品攒足了 分成 50份
先高浓度 IP down 整死那个 No. 50
再一步一步 浓度 减法 Immunoprecipitate 下一份 No.49
>No.48
.
.
.
>
>
>
>No. 01
也未尝不可
【在 s******s 的大作中提到】
: 这个多了，取决于你作用强不强，背景怎么样了。没具体做实验没人知道，你从弱的一
: 点点往上做吧。
h*2
16 楼
Let's pray for it!
J*n
17 楼
500W的电源能上双780Ti显卡SLI么？
【在 h**2 的大作中提到】
: 疯狂
: 花20-30块找个430-500w corsair或者ocz或者enermax，挺10年
【在 h**2 的大作中提到】
: 疯狂
: 花20-30块找个430-500w corsair或者ocz或者enermax，挺10年
e*f
18 楼
Good news, thanks.
a*o
19 楼
嗯非要装两个高端显卡的话这个电源是个好丢。
【在 J*********n 的大作中提到】
: 500W的电源能上双780Ti显卡SLI么？
【在 J*********n 的大作中提到】
: 500W的电源能上双780Ti显卡SLI么？
P*1
20 楼
３０１２其实对两党都没有什么政治益处，当然也没坏处。不同于非法移民，活活塞进
一批人，站在哪边就很重要了，利弊就很明显。３０１２就象路边的石头，他们就踢着
玩，个小，也不扎他们的脚，直到有一天，都不爱踢了，想换个踢踢，也就过了。觉得
那一天快到了。
一批人，站在哪边就很重要了，利弊就很明显。３０１２就象路边的石头，他们就踢着
玩，个小，也不扎他们的脚，直到有一天，都不爱踢了，想换个踢踢，也就过了。觉得
那一天快到了。
D*3
21 楼
$35 收了个 Antec EarthWatts EA-650
估计能推的动刚买的gtx460 2win
估计能推的动刚买的gtx460 2win
S*9
22 楼
淡定
【在 P****1 的大作中提到】
: ３０１２其实对两党都没有什么政治益处，当然也没坏处。不同于非法移民，活活塞进
: 一批人，站在哪边就很重要了，利弊就很明显。３０１２就象路边的石头，他们就踢着
: 玩，个小，也不扎他们的脚，直到有一天，都不爱踢了，想换个踢踢，也就过了。觉得
: 那一天快到了。
【在 P****1 的大作中提到】
: ３０１２其实对两党都没有什么政治益处，当然也没坏处。不同于非法移民，活活塞进
: 一批人，站在哪边就很重要了，利弊就很明显。３０１２就象路边的石头，他们就踢着
: 玩，个小，也不扎他们的脚，直到有一天，都不爱踢了，想换个踢踢，也就过了。觉得
: 那一天快到了。
x*u
23 楼
I hope so!!
【在 P****1 的大作中提到】
: ３０１２其实对两党都没有什么政治益处，当然也没坏处。不同于非法移民，活活塞进
: 一批人，站在哪边就很重要了，利弊就很明显。３０１２就象路边的石头，他们就踢着
: 玩，个小，也不扎他们的脚，直到有一天，都不爱踢了，想换个踢踢，也就过了。觉得
: 那一天快到了。
【在 P****1 的大作中提到】
: ３０１２其实对两党都没有什么政治益处，当然也没坏处。不同于非法移民，活活塞进
: 一批人，站在哪边就很重要了，利弊就很明显。３０１２就象路边的石头，他们就踢着
: 玩，个小，也不扎他们的脚，直到有一天，都不爱踢了，想换个踢踢，也就过了。觉得
: 那一天快到了。
n*e
24 楼
这样看来,IV前两天解释另外两个匿名hold,还是有点靠谱的.之前非常怀疑他们那个解
释.
释.
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