
Overview:
SMAD5 is a member of the SMAD family and mediates signaling by the transforming growth factor-beta (TGFB) superfamily and related ligands (1). SMAD5 plays a critical role in the signaling pathway by which TGFB inhibits the proliferation of human hematopoietic progenitor cells. SMAD5 is up-regulated in gastric epithelial cells during the infection of the pathogen Helicobacter pylori and it mediates apoptosis of gastric epithelial cells induced by H. pylori infection (2). In mature human B cells, bone morphogenetic protein 6 (BMP-6) inhibits cell growth and rapidly induces phosphorylation of SMADs 5 and 8.
Gene Aliases:
Dwfc, JV5-1, MADH5, DKFZp781C1895, DKFZp781O1323
Genbank Number:
NM_005903
References:
1. Heldin, C.H. et al: TGF-beta signalling from cell membrane to nucleus through SMAD proteins". Nature, 1997, 390 (6659): 465-71. 2. Nagasko, T. et al: Up-regulated Smad5 mediates apoptosis of gastric epithelial cells induced by Helicobacter pylori infection. J Biol Chem. 2003 Feb 14;278(7):4821-5.
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植物材料 用最精确的方法,称取不超过100mg的植物材料,置于处理过的研钵,加入液氮进行研磨 将研磨得到的粉末,快速转移至无RNase,并经过液氮冷却的2mL离心管(自备),
但是,这样做的前提的,RNA提得要好,浓度要高,这样免得RNA降解到不能用了。因为在有二价阳离子的(DNAseI buffer含有)情况下,RNA在60度以上时必然发生非酶促降解。
想搞清一个问题,如何区别是因为small RNA 引起的调控呢?还是tRNA引起的调控,有没有有效的分离这两种RNA的办法呢?
样品处理:
a. 植物组织:取新鲜或-70℃冻存100mg组织在液氮中研磨,把粉末加入到1ml裂解液中混匀。
b. 动物组织:取新鲜或-70℃冻存100mg组织加1ml裂解液,用组织研磨杵或匀浆器匀浆处理。
c. 贴壁细胞:直接在培养板中加入裂解液裂解细胞,每106细胞加1ml 裂解液。用取样器吹打混匀。

