
Overview:
SMAD3 is a direct mediator of transcriptional activation by the TGFB receptor. The activity of SMAD3 is regulated by the TGFB receptors, and SMAD3 is phosphorylated and associated with the ligand-bound receptor complex. TGFB stimulation leads to phosphorylation and activation of SMAD3, which form a complex with SMAD4 that accumulate in the nucleus and regulate transcription of target genes such as CDK inhibitor (1). SMAD3 containing a C-terminal truncation acts as a dominant-negative inhibitor of the normal TGFB response. SMAD3 is a major physiologic substrate of the G1 cyclin-dependent kinases CDK4 and CDK2 (2).
Gene Aliases:
MADH3, JV15-2, HSPC193, HsT17436, MGC60396, DKFZP586N0721, DKFZp686J10186
Genbank Number:
NM_005902
References:
1. Inman, G. J. et al: Nucleocytoplasmic shuttling of Smads 2, 3, and 4 permits sensing of TGF-beta receptor activity. Molec. Cell 10: 283-294, 2002. 2. Matsuura, I. et al: Cyclin-dependent kinases regulate the antiproliferative function of Smads. Nature 430: 226-231, 2004.
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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。

