GST pull down(GST融合蛋白沉降技术)是体外检测蛋白相互作用的常用方法,一般来说,GST融合蛋白pull down方法用于两个方面:一是鉴定能与已知融合蛋白相互作用的未知蛋白质;二是鉴定两个已知蛋白质之间是否存在相互作用。为排除假阳性所设的一组对照实验是在没有诱饵蛋白存在的条件下,将样品结合到GST上。该对照可以是表...
1.RNA Pull Down Assay(1) 为了鉴定特定的RNA分子是否与某一种特定的蛋白结合,研究人员根据DNA-Protein实验中DNA Pull down的实验方法进行改进,使其在RNA上实现。 技术步骤如下: (1)设计末端携带T7 RNA Promoter的引物扩增DNA片段; (2)利用T7 RNA polymerase 进行转录,转录过程添加携带生物素的dNTP,使转录本携...
Here we describe an RNA-protein pull-down assay, which enables the identification of proteins that interact with an RNA under study. As an example we describe pull-down of proteins interacting with ncRNA XIST, which assists in the recruitment of the polycomb-repressive complex-2 (PRC2) and ...
The control system for the pull-down assay uses 3? untranslated-region androgen receptor (AR) RNA, poly(A)25 RNA and mammalian cell lysate. The proximal 3? untranslated region (UTR) of AR RNA contains UC-rich regions for HuR and Poly(C) Binding Proteins (CP1 and CP2). These RNA-...
Noncoding RNAPull-downRNA-protein interactionsRecent advances in next-generation sequencing have revealed that majority of the plant genome is transcribed into long noncoding RNA (lncRNA). Many lncRNAs function by interacting with proteins and......
The control system for the pull-down assay utilizes 3'-untranslated region androgen receptor (AR) RNA, poly(A)25 RNA, and mammalian cell lysate. The proximal 3'-untranslated region (UTR) of androgen receptor RNA contains UC-rich regions for HuR and Poly(C) Binding Proteins (CP1 and 2)....
The full length p65 expression plasmid was transiently transfected into Hela cells and protein pull-down experiments were performed using anti-FLAG antibody conjugated to agarose A/G beads. Following the pull downs, total RNA was extracted from the bound protein fraction and reverse-transcription ...
RNA-Protein pull down 文档格式: .pdf 文档大小: 149.7K 文档页数: 8页 顶/踩数: 0/0 收藏人数: 0 评论次数: 0 文档热度: 文档分类: 待分类 文档标签: RNA-Proteinpulldown 系统标签: rnaproteinhurpiercelabelingutr INSTRUCTIONSPierceBiotechnologyPOBox117(815)968-0747.thermoscientific/pierce3747N.Meridia...
RNA pull-down assay GAS5 sense and antisense sequences, VAV1 coding region (CDS) sense and antisense sequences were in vitro transcribed from pGEM-T plasmid and biotinylated by Sangon Biotech (Shanghai) Co., Ltd. (Shanghai, China). Next, RNA pull down assay was performed in whole-cell ...
Cells were collected and lysed in IP lysis buffer (as per ‘MS2 pulldown assay’). The protein concentration was measured using the Bradford method. DHX15, p65 or p38 was immunoprecipitated after incubating the cell lysates with antibody for 6 h and an additional 2 h with Protein G Sep...