干细胞之家 - 中国干细胞行业门户第一站

 

 

搜索
中源协和

免疫细胞治疗专区

欢迎关注干细胞微信公众号

  
查看: 54023|回复: 0
go

求红色荧光蛋白基因(RFP),可以交换,我有YFP和CFP [复制链接]

Rank: 1

积分
0 
威望
0  
包包
61  
楼主
发表于 2010-5-12 19:01 |显示全部帖子
干细胞之家微信公众号
你好,我们实验室有m cherry这种蛋白,具体的我也没查过,但是可以发红光,是同学从法国带来的,他说这个就是GFP,你再查查看,如果你能用的话,就和我联系,qq:409478437。我也正好需要YFP和CFP,能否交换?
( a  L" {: w) G# }    以下资料你看看:Clontech is distributing vectors encoding several of the Living Colors® Fruit Fluorescent Proteins, including mCherry and tdTomato, which are already well characterized and recognized in the literature, and were developed in Dr. Roger Tsien’s lab (1–3). They have a wide emission range, three antibodies for detection, demonstrated stable expression, and perform successfully in numerous fusion applications. The Fruit Fluorescent Proteins are mutants derived from mRFP1, a monomeric mutant of DsRed, by directed mutagenesis (4). ) n% v: D4 i# X% w

1 t3 i  `, Z9 l2 yEffective Fusion Constructs
7 E3 ?! ^6 L/ W- X; j
1 ^3 H' }; G' |+ T7 B" W5 _mCherry has been successfully fused to several proteins, including actin and tubulin (Figure 1). Other fusion proteins containing mCherry have been reported in Arabidopsis (5), zebrafish (6), E. coli (7), HIV virions (8), and yeast (9). These fusions have also been used for quantitative imaging techniques including fluorescence resonance energy transfer (FRET; 10), fluorescence recovery after photobleaching (FRAP; 11), and fluorescence lifetime imaging microscopy (FLIM; 12).
2 |- \7 @/ l# Z! A1 g
- B5 G# l& T, A, D9 L( ?- tStable Cell Lines
/ a/ l- o# C  z+ [
' \0 q3 T6 e5 D" N* ^, a" G! LWe have established three stably-transfected HEK 293 cell lines with different levels of mCherry expression (as measured by flow cytometry). Transfected cells were observed to grow at a rate similar to nontransfected control cells, without increased cell death, as determined by visual inspection.! f1 X( g" E9 ?0 F% g8 w) S

$ O2 n3 H5 q% u! d2 [* N' bSimple Antibody Detection and Differentiation
9 S" ^% F5 z4 \( O/ \, C. R' r2 R: ?
3 s2 W' k. m2 n0 b% |  Q7 QUse the mCherry Monoclonal Antibody (Cat. No. 632543) to detect the mCherry, mOrange, mOrange2, mPlum, and mStrawberry Fruit Fluorescent Proteins by Western blot. Alternatively, the DsRed Monoclonal and Polyclonal Antibodies can be used to detect these Fruit Fluorescent Proteins.0 {* u' h7 j& c# v: }& X( Y% }  A5 Y' p
2 |( _  u+ h8 _  F8 Z3 n+ l- O
Use the DsRed Monoclonal and Polyclonal Antibodies (Cat. Nos. 632392, 632393, & 632496) to detect the mBanana, mRaspberry, and tdTomato Fruit Fluorescent Proteins by Western blot analysis (13).
‹ 上一主题|下一主题 ›
你需要登录后才可以回帖 登录 | 注册
验证问答 换一个

Archiver|干细胞之家 ( 吉ICP备2021004615号-3 )

GMT+8, 2026-11-10 07:51

Powered by Discuz! X1.5

© 2001-2010 Comsenz Inc.