RNAi Machinery Antibody Sampler Kit

• 产品编号：CST-4487S      品牌：Cell Signaling Technology       原厂货号：4487S
• 产品分类：抗体 > 一抗 > 复合抗体试剂盒
• 应用分类：

其它组分：

描述：

RNAi Machinery Antibody Sampler Kit中每一种抗体检测相应的内源靶蛋白水平，并且不与其它蛋白发生交叉反应。通过人工合成人类Dicer蛋白Ala1123位点或小鼠Mili蛋白Leu118位点周围相应的多肽片段去免疫动物从而制备多克隆抗体。通过蛋白A和多肽亲和层析纯化多克隆抗体。通过人工合成小鼠Argonaute 2蛋白或人类Drosha蛋白Gly953位点周围相应的多肽片段去免疫动物从而制备单克隆抗体。该RNAi Machinery Antibody Sampler Kit提供了分析内源性RNA干扰相关蛋白的一个经济的方式。该试剂盒含有进行免疫印迹（western blot）实验所需的一抗和二抗。

RNA干扰(RNAi)是真核生物中一个整体水平的基因调控机制。凭借与Dicer、Drosha, Argonaute 2 (Ago2)和Miwi/Mili蛋白相互作用，microRNA (miRNA)在细胞核内被加工，且被应用到基因沉默及下调基因表达。Dicer是RNase III酶家族成员之一，它能特异性剪切双链RNAs从而产生microRNAs (miRNAs) (1)。核内的III类RNase Drosha将长的初始转录本pri-miRNAs剪切成含有茎环pre-miRNAs之后(2)，pre-miRNAs被转运到细胞质，随后进一步被Dicer酶剪切成含22个核苷酸的成熟miRNAs(3)。然后，成熟miRNAs成为RNA诱导沉默复合物（RNA-Induced Silencing Complex，RISC）的一部分，从而能够结合目标mRNA的3'端非编码区(3)。Drosha酶的功能干扰后将导致pri-miRNAs的增加和pre-miRNAs的减少(2)。Drosha与它成分如DGCR8组成成为微加工器的多蛋白复合物的一部分(4)。Drosha与DGCR8蛋对miRNA生物合成是必需的(5)。Argonaute蛋白参与microRNA介导基因沉默的不同的阶段，例如翻译的抑制和mRNA降解(6)。果蝇piwi 基因被鉴定作为生殖系干细胞自我更新必需的，且在不同的物种包括拟南芥，秀丽隐杆线虫和人类中，Piwi的同源性被很好的保存下来(7)。Miwi和Mili蛋白是小鼠Piwi 同源物，都包含有羧基端Piwi结构域。该功能域能结合到在雄性生殖细胞中Piwi-interacting RNAs (piRNAs)，并且对小鼠精子的生成有着至关重要的作用(8-11)。

原厂资料：

Specificity / Sensitivity

Each antibody in the RNAi Machinery Antibody Sampler Kit recognizes endogenous levels of their target protein and does not cross-react with other proteins.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to the residues surrounding Ala1123 of human Dicer or residues surrounding Leu118 of mouse Mili protein. Polyclonal antibodies are purified by protein A and peptide affinity chromatography. Monoclonal antibody is produced by immunizing animals with synthetic peptides corresponding to the sequence of mouse Argonaute 2 or around Gly953 of human Drosha proteins.

Description

The RNAi Machinery Antibody Sampler Kit provides an economical means to analyze proteins associated with endogenous RNA interference. The kit contains enough primary and secondary antibodies to perform four western blot experiments.

Background

RNA interference (RNAi) serves as a global mechanism of gene regulation in eukaryotes. Through interactions with Dicer, Drosha, Argonaute 2 (Ago2) and Miwi/Mili proteins, microRNA (miRNA) is processed within the nucleus and utilized for gene silencing and down regulation of gene expression. Dicer is a member of the RNase III family that specifically cleaves double-stranded RNA to generate microRNA (miRNA) (1). Long, primary transcripts (pri-miRNAs) are processed to stem-looped pre-miRNAs by the nuclear RNase III Drosha (2) and are then transported to the cytoplasm for further processing by Dicer to produce mature, 22-nucleotide miRNAs (3). The mature miRNA then becomes a part of the RNA-Induced Silencing Complex (RISC) and can bind to the 3' UTR of the target mRNA (3). Interference of Drosha pri-miRNA processing results in the increase of pri-miRNAs and the decrease of pre-miRNAs (2). Drosha forms part of a multiprotein complex called the Microprocessor along with other components, such as DGCR8 (4). Both Drosha and DGCR8 are necessary for miRNA biogenesis (5). Argonaute protein family members participate in various steps of miRNA-mediated gene silencing such as repression of translation and mRNA turnover (6). The Drosophila piwi gene was identified as being required for the self-renewal of germ-line stem cells, and its homologues are well conserved among various species including Arabidopsis, C. elegans and human (7). Miwi and Mili proteins are both mouse homologs of Piwi and contain a carboxy-terminal Piwi domain that binds to Piwi-interacting RNAs (piRNAs) in male germ cells and are essential for spermatogenesis in mouse (8-11).

注意事项：

Storage: Supplied in 10 mM sodium HEPES (pH 7.5), 150
mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02%
sodium azide. Store at –20°C.
Do not aliquot the antibody