concentration and kinds of detergent for cell (293T) lysis?
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.
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!
【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
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
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!
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