
Overview:
NEK6 is a serine/threonine kinase that is a member of the Nek family of protein kinases that share an amino-terminal catalytic domain related to NIMA family (1). Nek6 is a nuclear and cytoplasmic kinase that is required for mitotic progression of human cells. Nek6 is phosphorylated and activated during M phase of the cell cycle (2). Inhibition of Nek6 function by either overexpression of an inactive Nek6 mutant or elimination of endogenous Nek6 by siRNA arrests cells in M phase and triggers apoptosis.
Gene Aliases:
SID6-1512
Genbank Number:
NM_014397
References:
1. Kandli,M. et al: Isolation and characterization of two evolutionarily conserved murine kinases (Nek6 and nek7) related to the fungal mitotic regulator, NIMA. Genomics. 2000 Sep 1;68(2):187-96.2. Yin, MJ. Et al: The serine/threonine kinase Nek6 is required for cell cycle progression through mitosis. J Biol Chem. 2003 Dec 26;278(52):52454-60.
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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。

