
Overview:
MLK3 or Mixed-Lineage Kinase 3 is a mitogen-activated protein kinase kinase kinase capable of preferentially activating MAPK8/JNK kinase and functions as a positive regulator of JNK signaling pathway (1). MLK3 can directly phosphorylate and activate IkappaB kinase alpha and beta, and is found to be involved in the transcription activity of NF-kappaB mediated by Rho family GTPases and CDC42. MLK3 is a signal-integrating kinase with conventional MAP3K catalytic activity and additional noncatalytic functions that contribute to RAF/ERK signalling (2). MLK3 is a component of the BRAF/RAF1 complex and is required for integrity of the complex and for activation of ERK by the complex.
Gene Aliases:
MAP3K11, PTK1, SPRK, MLK3, MGC17114, MLK-3
Genbank Number:
NM_002419
References:
1. Ing, Y. L. et al: MLK-3: identification of a widely-expressed protein kinase bearing an SH3 domain and a leucine zipper-basic region domain. Oncogene 9: 1745-1750, 1994. 2. Chadee, D. N. et al: MLK3 is required for mitogen activation of B-Raf, ERK and cell proliferation. Nature Cell Biol. 6: 770-776,2004.
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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。

