
Overview:
SGK3 is a member of the SGK family and encodes a phosphoprotein with a PX (phox homology) domain. PKD1 can phosphorylate and activate SGK3 in vitro (1). A 10-fold increase in PKD1 increases the phosphorylation status of SGK3. When expressed in human embryonic kidney cells, IGF1 and peroxide significantly activate SGK3, and the activation could be reduced by preincubation with inhibitors of PI3 kinase. A yeast 2-hybrid screen found direct interaction between human SGK3 and GSK3?. SGK3 phosphorylates several target proteins and has a role in neutral amino acid transport and activation of potassium and chloride channels (2).
Gene Aliases:
CISK, SGKL
Genbank Number:
NM_013257
References:
1. Kobayashi, T. et al: Characterization of the structure and regulation of two novel isoforms of serum- and glucocorticoid-induced protein kinase. Biochem. J. 344: 189-197, 1999. 2. Gamper, N. et al: K+ channel activation by all three isoforms of serum- and glucocorticoid-dependent protein kinase SGK. Europ. J. Physiol. 445: 60-66, 2002.
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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。

