24小时热门版块排行榜

返回列表

当前只显示满足指定条件的回帖，点击这里查看本话题的所有回帖

康洁

新虫 (正式写手)

应助: 0 (幼儿园)
金币: 2.9
散金: 11
红花: 3
帖子: 330
在线: 104.4小时
虫号: 2006822
注册: 2012-09-17
专业: 配位化学

[交流] 谁有用荧光探针测过氧化氢的文献（最好是生物体内）

谁有用荧光探针测过氧化氢的文献（最好是生物体内）

回复此楼

» 猜你喜欢

1楼2014-04-24 22:40:44

已阅回复此楼关注TA 给TA发消息送TA红花 TA的回帖

brilliant

捐助贵宾 (职业作家)

应助: 118 (高中生)
金币: 12307.3
散金: 262
红花: 47
帖子: 4907
在线: 1107.3小时
虫号: 71523
注册: 2005-05-31
性别: GG
专业: 配位化学

★ ★ ★ ★ ★ ★ ★
小木虫: 金币+0.5, 给个红包，谢谢回帖
黄酮: 金币+1, 谢谢回帖交流 2014-04-29 09:19:52
康洁(likedong1021代发): 金币+5, 感谢回帖交流！ 2014-07-28 04:51:19

Nature Protocols | Protocol
PrintEmailShare/bookmark
Cite U LikeFacebookTwitterDeliciousDiggGoogle+LinkedInRedditStumbleUponPrevious articleNature Protocols | Protocol
Monitoring in vivo reversible cysteine oxidation in proteins using ICAT and mass spectrometry Next articleNature Protocols | Protocol
In vivo introduction of transgenes into mouse sciatic nerve cells in situ using viral vectors

Synthesis, labeling and bioanalytical applications of a tris(2,2′-bipyridyl)ruthenium(II)-based electrochemiluminescence probe
Xiaoming Zhou,1, Debin Zhu,1, Yuhui Liao,1, Weipeng Liu,1, Hongxing Liu,1, Zhaokui Ma1, & Da Xing1,
AffiliationsContributionsCorresponding author Journal name:
Nature Protocols
Volume:
9,
Pages:
1146–1159
Year published:
(2014)DOI:
doi:10.1038/nprot.2014.060
Published online17 April 2014 Article toolsFull text

PDF
Synthesis, labeling and bioanalytical applications of a tris(2,2′-bipyridyl)ruthenium(II)-based electrochemiluminescence prob...
Download as PDF (921 KB) View interactive PDF in ReadCube CitationReprintsRights & permissionsArticle metrics
Abstract
Abstract• References• Author information• Supplementary information Assays using probes labeled with electrochemiluminescent moieties are extremely powerful analytical tools that are used in fields such as medical diagnostics, environmental analysis and food safety monitoring, in which sensitive, reliable and reproducible detection of biomolecules is a requirement. The most efficient electrochemiluminescence (ECL) reaction to date is based on tris(2,2′-bipyridyl)ruthenium(II) (Ru(bpy)32+) with tripropylamine (TPrA) as the co-reactant. Here we present a detailed protocol for preparing Ru(bpy)32+ probes and their bioanalytical applications. This protocol includes (i) the synthesis of a biologically active Ru(bpy)32+-N-hydroxysuccinimide (NHS) ester, (ii) its covalent labeling with both antibodies and DNA probes and (iii) the detection and quantification of ECL in a microfluidic system with a paramagnetic microbead solid support. In our magnetic bead–based ECL system, two probes are required: a capture probe (labeled with biotin to be captured by a streptavidin-coated magnetic bead) and a detector probe (labeled with Ru(bpy)32+). The complex consisting of the analyte, the capture probe, the detector probe and the magnetic bead is brought into contact with the electrode by using a magnetic field. The Ru(bpy)32+ reacts with TPrA in solution to generate the ECL signal. The full protocol, including the synthesis and labeling of the bioactive Ru(bpy)32+, requires 5–6 d to complete. ECL immunoassays or nucleic acid tests only require 1.5–2 h, including the sample preparation time.